Modified proteins, isolated novel peptides,and uses thereof

ABSTRACT

The present invention provides several modified ABC transporter polypeptides that exhibit novel localization in the plasma membrane of polarized and non-polarized cells. The modified ABC transporter of the invention comprises the amino acid sequence of a native apically targeted ABC transporter, in particular cMOAT, MDR3 or MRP4, wherein the terminal tripeptide T-K-F motif of said native ABC transporter is mutated. The isolated modified ABC transporter polypeptide of the invention, and the nucleotide sequence encoding said polypeptide, have utility in the following applications: First, they are used to induce a drug resistant phenotype in a cell. Second, they are used to protect non-polarized cells during chemotherapy and other therapeutic applications. Third, they are used to produce novel cell lines that are used to screen for novel agonists or antagonists of the corresponding native ABC transporter polypeptides.

FIELD OF THE INVENTION

[0001] The present invention relates generally to novel proteins thatare capable of modulating the drug resistance of cells, tissues, organsand whole organisms. More specifically, the present invention providesseveral modified forms of ATP-Binding Cassette transporter (hereinafter“ABC pump” or “ABC transporter”) polypeptides that are normallylocalized in the canalicular (apical) membrane of polarized cells wherethey modulate the transport or efflux of one or more drugs, antibiotics,or other chemical compounds, wherein the modified ABC transporters ofthe invention are localized in the basolateral membrane of polarizedcells, or accumulate in the plasma membrane of a non-polarized cell.Several modified canalicular multispecific organic anion transporter(cMOAT) polypeptides (also known in the art as “MRP2”), a modified MDR3polypeptide, and a modified MRP4 polypeptide are exemplified herein thatare capable of being differentially expressed or localized within thecell membrane compared to the non-modified form of said polypeptides.The modified ABC transporter polypeptides of the invention are furthercapable of modulating the resistance of cells to a range of compounds,including antibiotics, chemotherapeutic agents, and antifungalcompounds, and, accordingly, the present invention clearly extends tothe uses of both the isolated modified ABC transporter polypeptide ofthe invention and the nucleotide sequence encoding same to: (i) induce amultidrug resistant phenotype in a cell; and (ii) protect polarized andnon-polarized cells during chemotherapy and other applications. Themodified ABC transporter polypeptides of the invention are alsoparticularly useful in screening for compounds that modulate theactivity (i.e. agonists or antagonists) of an ABC transporterpolypeptide, or to determine if the efflux of a particular compound ismodulated by a specific ABC transporter polypeptide. High throughputscreening protocols are described herein. The present invention furtherprovides isolated nucleic acids encoding the modified ABC transporterpolypeptide and gene constructs comprising same.

[0002] General

[0003] This specification contains nucleotide and amino acid sequenceinformation prepared using the programme PatentIn Version 3.1, presentedherein after the bibliography. Each nucleotide or amino acid sequence isidentified in the sequence listing by the numeric indicator <210>followed by the sequence identifier (e.g. <210>1, <210>2, etc). Thelength, type of sequence (DNA, protein (PRT), etc) and source organismfor each nucleotide or amino acid sequence are indicated by informationprovided in the numeric indicator fields <211>, <212> and <213>,respectively. Nucleotide and amino acid sequences referred to in thespecification are defined by descriptor “SEQ ID NO:” followed by thenumeric identifier. For example, SEQ ID NO: 1 refers to the informationprovided in the numeric indicator field designated <400>1, etc.

[0004] For the purposes of nomenclature, the nucleotide sequence of thenative cMOAT-encoding gene of humans is set forth in SEQ ID NO: 1, andthe corresponding amino acid sequence is set forth in SEQ ID NO: 2. TheC-terminal portion of native cMOAT is also presented in SEQ ID NO: 37.

[0005] The nucleotide sequence of a first modified cMOAT-encoding geneis set forth in SEQ ID NO: 3, and the corresponding amino acid sequenceis set forth in SEQ ID NO: 4. The amino acid sequence of SEQ ID NO: 4corresponds to the ΔcMOAT polypeptide of the invention (also termedherein “ΔT1543 ΔK1544 ΔF1545”), the C-terminal portion of which ispresented in SEQ ID NO: 44.

[0006] The nucleotide sequence of a second modified cMOAT-encoding geneis set forth in SEQ ID NO: 5, and the corresponding amino acid sequenceis set forth in SEQ ID NO: 6. The amino acid sequence of SEQ ID NO: 6corresponds to the T1543A K1544P F1545V polypeptide of the invention,the C-terminal portion of which is presented in SEQ ID NO: 38.

[0007] The nucleotide sequence of a third modified cMOAT-encoding geneis set forth in SEQ ID NO: 7, and the corresponding amino acid sequenceis set forth in SEQ ID NO: 8. The amino acid sequence of SEQ ID NO: 8corresponds to the S1542A polypeptide of the invention, the C-terminalportion of which is presented in SEQ ID NO: 39.

[0008] The nucleotide sequence of a fourth modified cMOAT-encoding geneis set forth in SEQ ID NO: 9, and the corresponding amino acid sequenceis set forth in SEQ ID NO: 10. The amino acid sequence of SEQ ID NO: 10corresponds to the T1543A polypeptide of the invention, the C-terminalportion of which is presented in SEQ ID NO: 40.

[0009] The nucleotide sequence of a fifth modified cMOAT-encoding geneis set forth in SEQ ID NO: 11, and the corresponding amino acid sequenceis set forth in SEQ ID NO: 12. The amino acid sequence of SEQ ID NO: 12corresponds to the K1544A polypeptide of the invention, the C-terminalportion of which is presented in SEQ ID NO: 41.

[0010] The nucleotide sequence of a sixth modified cMOAT-encoding geneis set forth in SEQ ID NO: 13, and the corresponding amino acid sequenceis set forth in SEQ ID NO: 14. The amino acid sequence of SEQ ID NO: 14corresponds to the F1545A polypeptide of the invention, the C-terminalportion of which is presented in SEQ ID NO: 42.

[0011] The nucleotide sequence of a seventh modified cMOAT-encoding geneis set forth in SEQ ID NO: 15, and the corresponding amino acid sequenceis set forth in SEQ ID NO: 16. The amino acid sequence of SEQ ID NO: 16corresponds to the T1543A K1544A F1545A polypeptide of the invention,the C-terminal portion of which is presented in SEQ ID NO: 43.

[0012] The nucleotide sequence of a modified MDR3-encoding gene is setforth in SEQ ID NO: 48, and the corresponding amino acid sequence of amodified human MDR3 polypeptide of the invention, lacking the T-K-Fmotif (i.e. the terminal 4 amino acids have been deleted), is presentedin SEQ ID NO: 49. The modified MDR3-encoding sequence is amplified fromnative human MDR3 cDNA using the primer sequences set forth in SEQ IDNO: 59 and SEQ ID NO: 60.

[0013] The nucleotide sequence of a modified MRP4-encoding gene is setforth in SEQ ID NO: 50, and the corresponding amino acid sequence of amodified human. MRP4 polypeptide of the invention, lacking the T-K-Fmotif (i.e. the terminal 3 amino acids have been deleted), is presentedin SEQ ID NO: 51. The modified MRP4-encoding sequence is amplified fromnative human MRP4 cDNA using the primer sequences set forth in SEQ IDNO: 61 and SEQ ID NO: 62.

[0014] The designation of nucleotide residues referred to herein arethose recommended by the IUPAC-IUB Biochemical Nomenclature Commission,wherein A represents Adenine, C represents Cytosine, G representsGuanine, T represents thymine, Y represents a pyrimidine residue, Rrepresents a purine residue, M represents Adenine or Cytosine, Krepresents Guanine or Thymine, S represents Guanine or Cytosine, Wrepresents Adenine or Thymine, H represents a nucleotide other thanGuanine, B represents a nucleotide other than Adenine, V represents anucleotide other than Thymine, D represents a nucleotide other thanCytosine and N represents any nucleotide residue.

[0015] Reference herein to prior art, including any one or more priorart documents, is not to be taken as an acknowledgment, or suggestion,that said prior art is common general knowledge in Australia or forms apart of the common general knowledge in Australia.

[0016] Throughout this specification, unless the context requiresotherwise, the word “comprise”, or variations such as “comprises” or“comprising”, will be understood to imply the inclusion of a stated stepor element or integer or group of steps or elements or integers but notthe exclusion of any other step or element or integer or group ofelements or integers.

[0017] As used herein the term “derived from” shall be taken to indicatethat a specified integer may be obtained from a particular source alb itnot necessarily directly from that source.

[0018] Those skilled in the art will appreciate that the inventiondescribed herein is susceptible to variations and modifications otherthan those specifically described. It is to be understood that theinvention includes all such variations and modifications. The inventionalso includes all of the steps, features, compositions and compoundsreferred to or indicated in this specification, individually orcollectively, and any and all combinations or any two or more of saidsteps or features.

[0019] The present invention is not to be limited in scope by thespecific embodiments described herein, which are intended for thepurposes of exemplification only. Functionally-equivalent products,compositions and methods are clearly within the scope of the invention,as described herein.

BACKGROUND TO THE INVENTION

[0020] The treatment of bacterial infections, fungal infections, and,more specifically, the treatment of various tumors and/or cancers, ofteninvolves the administration of chemotoxins and/or chemostatic compounds,which either kill or inhibit the growth of a tumor, such as, forexample, various anti-cancer chemotherapeutic agents, including vincaalkaloids, cisplatin, busulphan (busulfan), vincristine sulphate,merchlorethane, etoposide; and the administration of various chemicalcompounds which kill a host cell and/or invading pathogens, such as forexample, various antibiotic compounds. The majority of cytotoxic drugsare more effective against cells that are rapidly moving through thecell cycle, such as, for example, bacteria that are not in stationary orplateau phases, or tumors having a large growth fraction.

[0021] Transport of a drug to a tumor cell (i.e. influx), and itssubsequent efflux, are important factors governing the efficacy of anypharmaceutical agent, including a chemotherapeutic agent.

[0022] Pharmacokinetic and/or biochemical resistance, including, forexample, multidrug resistance (MDR) or pleiotropic resistance, to anadministered drug may occur over time. By “resistance”, is meant theability of a cell, group of cells, tissue, organ, or organism, to remainviable, grow or proliferate in the presence of a chemical compound.Accordingly, a “resistant” cell has the capacity to remain viable, andpreferably, to grow and/or to proliferate in the presence of saidchemical compound.

[0023] Hereinafter, the term “drug resistance” shall be taken to meanpharmacokinetic resistance and/or biochemical resistance, including thephenomenon of MDR, unless specifically stated otherwise.

[0024] Drug resistance is generally associated with a low concentrationof drug in the target cell, tissue, organ or organism. This is becauseof decreased intracellular accumulation of the drug, and/or defectivetransport, and/or reduced absorption, and/or altered drug distribution,and/or biotransformation of the drug, and/or enhanced elimination of thedrug from the site of administration and/or effect. The occurrence ofdrug resistance is one of the major obstacles to the successfultreatment of many conditions in humans and animals with such chemotoxinsand/or chemostatic compounds, such as, for example, various antibiotics,anti-fungal compounds, anti-viral compounds, and chemotherapeuticagents, in particular, the anthracyclines, epipodophyllotoxins, andvinca alkaloids.

[0025] In the case of anti-cancer treatments, drug resistance results ina sensitive tumor being converted into a resistant tumor that no longerresponds to chemotherapy.

[0026] On the other hand, it is also desirous to protect certain cells,tissues, and organs from cytostatic and/or cytotoxic compounds duringtreatment of disease, such as, for example, cancer or infection bypathogenic agents. The acquisition of drug resistance in those cells,tissues and organs is a highly desirable outcome. For example,protection of the haematopoietic system during chemotherapy may increasethe chances of survival of some cancer patients. Naturally, in suchcases it is important to maximize the efflux of the drug from the cellsin respect of which protection is sought.

[0027] Accordingly, drug resistance can hinder effective treatment.

[0028] Various drug resistant cell lines have been identified, such as,for example, rodent cell lines resistant to multiple drugs, andmultidrug resistant (MDR) lines derived from the human KB carcinoma cellline (a HeLa subclone). These lines were selected for their resistanceto colchicine, vinblastine, or adriamycin (see, for example, Kartner etal. (1983) Science 221,1285-1288; Akiyama et al. (1985) Somatic CellMol. Genet. 11, 117-126; Shen et al. (1986) J. Bio. Chem. 261,7762-7770; Shen et al. (1986) Science, 232: 643-645; and Shen et al.,(1986) Mol. Cell. Biol. 6, 4039-4044).

[0029] Efflux of a drug through the plasma membrane, which contributesto resistance of the cell, is mediated by one or more specific membranetransporters (Cole and Deeley (1998) Bioessays 20, 931-940; Goftesman etal (1995) Ann. Rev. Genet. 29, 607-649; Higgins et al (1992) Ann. Rev.Cell Biol. 8, 67-113). These membrane transporters belong to theso-called superfamily of ATP-Binding Cassette (ABC) transporters.

[0030] Cultured or primary epithelial cells, such as, for example,hepatocytes, neuronal cells, and certain cells of the immune system,maintain a characteristic polarized phenotype. The majority of plasmaproteins are distinguishable on the basis of their distribution eitherto the apical (canalicular) or to the basolateral membrane domain ofcultured or primary epithelial cells. Relatively few proteins have beenidentified that are equally distributed on both membrane domain surfacesof these cells (Mellman et al., (1993) J. Cell Sci., Suppl. 17, 1-7).The ABC transporters are generally targeted to the basolateral membraneof such polarized cells (e.g. MRP1, MRP3, and MRP6). Alternatively, ABCtransporters may be targeted to the apical (canalicular) membrane [e.g.the canalicular multispecific organic anion transporter (cMOAT) alsoknown as the multidrug resistance-associated protein 2 (MRP2), theP-glycoprotein (P-gp) transporter and its homologues (e.g. MDR2, MDR3),and MRP4].

[0031] Substrates for ABC transporters tend to be amphiphilic organiccations and anions.

[0032] Those skilled in the art will be aware that ABC transporters areresponsible for the transport of a wide range of compounds, such as, forexample, 4-NQO, sorbic acid, ketoconazole, econazole, oligomycin,antimycin, paromomycin, colchicine, vinblastine, and adriamycin.

[0033] It has also been shown that many pathogenic microorganisms, suchas Candida albicans and Plasmodium falciparum, can use the ABCtransporter-mediated drug efflux mechanism to evade the toxicity of anadministered therapeutic agent (Cowrnan and Foote (1990) Int. J.Parasitol. 20, 503-513; Foote et al (1989) Cell 57, 921-930; Prasad etal. (1995) Curr. Genet 27, 320-329; Sanglard et al (1995) Antimicrob.Agents Chemother. 39, 2378-2386), or to otherwise develop resistance.

[0034] The P-gp, MRP1, MRP2 (cMOAT), MRP3, MRP4, MRP5, MRP6, MDR2, andMDR3 proteins are all membrane-localized proteins that pump drugs out ofcells by an energy-dependent mechanism requiring ATP. In the liver,P-gp, MRP2 (cMOAT), MRP4, and MDR3, at least, transport a range oforganic compounds across the apical (canalicular) membrane into bile. Innon-polarized cells, MRP2 (cMOAT) accumulates in intracellular vesicles,with little accumulation of this protein in the plasma membrane (Harriset al., (2001) J. Biol. Chem 24, 20876-20881). In contrast to theseso-called “apical” ABC transporter proteins, the MRP1, MRP3, and MRP6proteins normally function in the basolateral (sinusoidal) membrane ofpolarized cells. Increased activity of the ABC transporters may lowerthe intracellular accumulation of a particular drug in all cells inwhich they are expressed, and result in the cell becoming resistant tothe administered drug.

[0035] Several human P-glycoproteins (P-gp), such as, for example, MDR1,MDR2, and MDR3, have been identified. The genes encoding these proteinsare homologous to the hamster mdr gene (see, for example, Roninson etal. (1984) Nature, 309, 626-628; Gros et al., (1986) Nature 323, 728-731and Gros et al. (1986) Proc. Natl. Acad Sci. USA, 83, 337-341). The MDR1and MDR2 proteins are expressed in multidrug-resistant human KBcarcinoma cell lines (Fojo et al., (1985) Proc. Natl. Acad. Sci. USA 82,7661-7665; Roninson et al (1986) Proc. Natl. Sci. USA 83, 4538-4542).The MDR1 gene encodes a 4.5-kb mRNA which is over expressed in all ofthe highly drug-resistant cell lines (Roninson et al. (1986) Proc. Natl.Sci. USA 83, 4538-4542; Shen et al. (1986) J. Bio. Chem. 261, 7762-7770;Shen et al. (1986) Science, 232: 643-645; and Shen et al., (1986) Mol.Cell. Biol. 6, 4039-4044), and in certain normal and tumor tissues (Fojoet al., (1987) Proc. Natl. Acad. Sci. USA 84, 265-269). Moreover, Chenet al., (1987) Cell 47, 381-389 have described the isolation of a set ofoverlapping cDNAs for the entire coding region of the human MDR1 mRNA.The human MDR1 gene is also expressed at high levels in murine cellsthat have been transformed to a drug resistant phenotype using genomicDNA derived from drug resistant human cells. This finding suggests thatexpression of the MDR1 gene can contribute to the development of the MDRphenotype (Shen et al (1986) J. Bio. Chem. 261, 7762-7770; Shen et al.(1986) Science, 232: 643-645; and Shen et al., (1986) Mol. Cell. Biol.6, 4039-4044). Several additional workers have demonstrated the abilityof an isolated, and expressed, murine mdr gene to confer multidrugresistance on isolated cells (Gros et al. (1986) Proc. Natl. Acad. Sci.USA, 83, 337-341; Gros et al., (1986) Nature 323, 728-731; Ueda et al.(1987) J. Biol. Chem. 262, 505-508).

[0036] Native P-glycoprotein is absent from most normal tissues, but avariety of tissues in mammals have been found to express P-gp in aninducible form, such as, for example, the kidney, liver, smallintestine, colon, uterine secretory epithelium, and adrenal gland. Inpolarized cells, such as those in the renal tubule or small intestinalmucosa, liver, and pancreas, P-gp is expressed in a polarized manner andis located in the luminal brush borders. Thus, P-gp is located on theapical surface of proximal tubule cells in the kidney, on the apicalsurface of intestinal epithelial cells, on the apical surface of smallductules of the pancreas and on the binary face of hepatocytes. Only inadrenal cells is P-gp is uniformly distributed in the membrane. Thenormal function of P-gp is not firmly established, but it is known thatit can remove toxic substances from cells (Gatmaitan and Arias (1993)Adv Pharmacol 24:77-97).

[0037] P-gp is phosphorylated in vivo, and early studies havedemonstrated that a change in the state of phosphorylation of P-gp hasbeen associated with differences in relative drug resistance ofmammalian cells, suggesting that the phosphorylation mechanisms may beinvolved in the regulation of the efflux activity of the drugtransporter (Center (1983) Biochem. Biophys. Res. Comm. 115, 159-166;Hamada et al (1987) Cancer Res. 47, 2860-2865).

[0038] P-gp-mediated drug resistance may be ameliorated to some extentvia the administration of P-gp modulators or antagonists that inhibitthe export function of P-gp, thereby allowing the accumulation of achemotherapeutic agent administered to the patient. However, P-gpmodulators are not useful in combination therapy for the simultaneousprotection of the haematopoietic system and anti-cancer treatment of thepatient, particularly where MDR1 is ectopically expressed inhaematopoietic cells and chemotherapeutic agents and P-gp modulators areadministered to inhibit or prevent tumorigenesis. This is because theP-gp modulator inhibits the activity of ectopically expressed MDR1protein, in addition to inhibiting the endogenous P-gp activity.

[0039] The cMOAT transporter activity was initially characterized inhepatocytes, by comparing normal rats to mutants (TR/GY) that lackedcanalicular transport activity (Oude Elferink, et aL (1995) BiochimBiophys Acta. 1241, 215-268). Evers, R., et al. (J Clin Invest. (1988)101, 1310-1319) demonstrated that the drug export activity ofrecombinant cMOAT protein in polarized kidney MDCK cells expressing acMOAT-encoding cDNA was confined predominantly to the apical membrane.Subsequently, immunostaining also revealed that the cMOAT protein ispredominantly expressed in the apical membrane of hepatocytes (Konig etal. (1999) Biochim Biophys Acta. 1461, 377-394).

[0040] The present inventors have also discovered that native cMOATfails to accumulate in the plasma membrane of non-polarized cells(Harris et al., (2001) J. Biol. Chem 24, 20876-20881). Based upon thisexpression pattern, the native cMOAT polypeptide is of limited utilityin conferring drug resistance on non-polarized cells, such as, forexample, certain cells of the haematopoietic system.

[0041] Native MRP1 transporter activity is enhanced in tumors exposed tochemotherapeutic agents, thereby conferring acquired resistance on thetumor cells (Goldstein, et al. (1989) J Natl Cancer Inst. 81,116-124;Slapak, C. A., et al. (1994) Blood 84, 3113-3121).

[0042] Phosphorylation has been proposed to regulate activity of the S.cerevislae Ycf1 protein. Ycf1 is an orthologue of human MRP1 located onthe vacuolar membrane of yeast cells (Li et al (1998) J. Biol. Chem.273, 33449-33454; Szczpka et al (1994) J. Biol. Chem. 269, 22853-22857).

[0043] For the treatment of some cancerous cells, such as those ofnon-small cell lung cancers, MRP1-mediated drug resistance in respect ofchemotherapeutic agents is not acquired. Rather, resistance occurs fromthe outset of treatment (i.e. intrinsic resistance), indicating a highconstitutive level of expression of the MRP1 protein (Zaman; G. J., etal., (1993) Cancer Res. 53, 1747-1750). As a consequence, the treatmentof patients having advanced tumors, relapsed tumors, or tumors whichexhibit intrinsic MRP1-mediated resistance, often requires high doses ofchemotherapeutic agent(s). The potential benefits of such high-dosageregimens are generally offset or compromised by myelosuppression,involving the destruction of bone marrow cells, that is induced by thecytotoxic chemotherapeutic agent used.

[0044] Protection of bone marrow from the cytotoxic effects ofchemotherapeutic agents has been attempted in murine models. Forexample, MDR1 has been expressed ectopically in murine bone marrowcells. However, in such procedures, myeloproliferative syndrome developsin the mice, wherein cells of certain haematopoietic lineagesdifferentiate and proliferate abnormally (Bunting, K. D., et al., (1998)Blood 92, 2269-2279).

SUMMARY OF THE INVENTION

[0045] In work leading up to the present invention, the inventors soughtto identify novel means for modulating the drug resistance of cellsmediated by ABC transporter polypeptides, so as to provide for improvedtreatment regimes and/or to reduce the adverse side-effects of drugs onthe haematopoietic system. The inventors have produced a modified ABCtransporter polypeptide having novel distribution characteristics in theplasma membrane of polarized and non-polarized cells. These noveldistribution characteristics facilitate the treatment of cells by genetherapy regimes, including the use of combination therapies involvingboth gene technology and traditional drug administration regimes.

[0046] More particularly, the present invention provides modified ABCtransporter polypeptides, including modified cMOAT, MDR3 and MRP4polypeptides, that are capable of being predominantly translocated tothe basolateral (sinusoidal) membrane, or localized in the plasmamembrane of a non-polarized cell. The modified polypeptide thus exhibitsa surprising and novel accumulation relative to the corresponding nativeABC transporter polypeptide.

[0047] Preferably, the modified ABC transporter polypeptide of thepresent invention consists of an active ABC transporter polypeptidecomprising a mutation wherein at least one amino acid residue in theC-terminal region of said active ABC transporter polypeptide issubstituted or deleted.

[0048] In an alternative embodiment, the modified ABC transporterpolypeptide of the present invention consists of an active ABCtransporter polypeptide comprising a mutation wherein at least one aminoacid residue of a tripeptide T-K-F motif present in said active ABCtransporter polypeptide is substituted or deleted.

[0049] The novel localization patterns for the modified ABC transporterpolypeptides described herein facilitates the efflux of certain liganddrugs from the cell to confer resistance properties thereon. The presentinvention clearly extends to any and all uses of the novel modified ABCtransporter polypeptides as described herein consistent with theirstated modes of action.

[0050] In particular, the modified ABC transporter polypeptide confersresistance to one or more chemical compounds on a cell. For example,resistance is conferred to a cytostatic or cytotoxic compound used inthe treatment of infection or disease. More particularly, the modifiedpolypeptides are useful when protection of non-polarized cells (e.g.cells of the haematopoietic system) is required during the treatment ofpatients with cytotoxic or cytostatic compounds.

[0051] In fact, the modified ABC transporter of the invention is usefulfor conferring resistance against any pharmaceutical agent that that ismetabolized by ABC transporters that are normally apically-localized inpolarized cells, by facilitating efflux through the basolateralmembrane.

[0052] Alternatively, or in addition, the modified ABC transporter ofthe invention is useful for conferring de novo resistance on anon-polarized cell by facilitating efflux through the plasma membrane.In a particularly preferred embodiment exemplified herein, resistance toBusulfan is conferred on L1210 cells by ectopically expressing amodified cMOAT polypeptide therein.

[0053] As a consequence of this conferred resistance, the modified ABCtransporter can be used to identify any potentially toxic agents at anearly stage, by screening chemical libraries, thereby identifying novelcytotoxins that would not otherwise be identified prior to clinicaltrials or use. Once identified, the correct dosage level of anypharmaceutical compound for a particular cell type or geneticbackground, to achieve a desired effect (e.g. toxicity) is readilydetermined.

[0054] Additionally, the modified ABC transporter polypeptide of theinvention is used in combination with modulators of heterologous ABCtransporters.

[0055] Additionally, the modified ABC transporter polypeptide of theinvention is used to develop novel cell lines for assaying ABCtransporter activity, substrate specificity, drug metabolism, or drugtransport.

[0056] The assays supra are particularly amenable to identifying newpharmaceuticals that modulate ABC transporter activity. Accordingly, afurther aspect of the invention contemplates a simple and reliable invivo screening system for the discovery of novel agonists andantagonists of an ABC transporter polypeptide. Additionally thescreening system can be used to determine if efflux by a certain ABCtransporter is a significant pathway in the metabolism of a particulardrug.

[0057] A further aspect of the present invention provides a geneconstruct comprising a nucleotide sequence encoding the modified ABCtransporter polypeptide of the invention. Preferably, the nucleic acidmolecule is operably linked to a promoter sequence to facilitate itsexpression in a bacterial cell, yeast, fungal cell, insect cell, ormammalian cell.

[0058] The gene construct according to this embodiment of the inventionis particularly useful for conferring novel drug resistancecharacteristics on a cell, in particular a non-polarized cell, oralternatively, for transporting particular drugs from the cell.Accordingly, a further aspect of the invention provides a cellcomprising the subject gene construct and preferably, which expressesthe modified ABC transporter polypeptide of the invention.

[0059] In a particularly preferred embodiment, non-polarized cells,(e.g. fibroblasts or cells of the haemopoietic system) are produced thatexpress the modified ABC transporter polypeptide generally within theplasma membrane where it functions in the efflux of certain ligand drugsfrom the cell to confer resistance properties thereon.

[0060] Alternatively, polarized cells, (e.g. cultured epithelial cellssuch as MDCK or Caco-2 cells, or primary epithelial cells such ashepatocytes, intestinal cells, or hippocampal neurons) are produced thatexpress the modified ABC transporter polypeptide predominantly in thebasolateral membrane.

[0061] A further aspect of the invention contemplates a transport signalpeptide to facilitate the efficient translocation or transcytosis of apolypeptide to the apical membrane of a polarized cell.

BRIEF DESCRIPTION OF THE DRAWINGS

[0062]FIG. 1 is a copy of a photographic representation of arepresentative Madine-Darby canine kidney (MDCK) cell expressingcMOAT-gfp in a confluent monolayer of cells. Fluorescence is evidentthroughout the cell in the top down view (upper panel). However, incross-section, the XZ view reveals specific apical (AP) localization andminimal basolateral (BL) targeting of protein (lower panel). Thecover-slip is detected as a line on the apical surface of the cells dueto autofluorescence. All scale bars indicate 5 microns.

[0063]FIG. 2 is a copy of a photographic representation showing thatconfluent MDCK cells expressing MRP1-gfp have a ringed appearance in thetop down view (upper panel) due to fluorescence in the basolateral (BL)membrane. In the XZ view (lower panel) the lateral targeting of MRP1-gfpis confirmed with the cell to cell membranes being defined. AP, apical.

[0064]FIG. 3 is a copy of a photographic representation showing thatconfluent MDCK cells expressing ΔcMOAT appear ringed in the top downview (upper panel) with a similar appearance to MRP1-gfp (FIG. 2). Inthe XZ view (lower panel), ΔcMOAT-gfp shows definite laterallocalization with the cell to cell membrane outlined by fluorescingprotein. Apical (AP) targeting is minimal compared with native cMOATfused to GFP (FIG. 1). BL, basolateral.

[0065]FIG. 4 is a copy of a photographic representation showing thelocalization of modified cMOAT-gfp fusion proteins comprising mutationsof the T-K-F motif of the cMOAT portion. Upper panels in each figurerepresent the top down view of the cells, whilst the lower panelsrepresent the XZ view of cells, as follows.:

[0066]FIG. 4A is a copy of a photographic representation showing thatthe T1543A mutant has a non-polarized distribution of the fusionprotein. Fluorescence was detected in both the apical (AP) andbasolateral (BL) membranes giving a ringed appearance from the top downview (upper panel), but the XZ view (lower panel) reveals the nonpolarized distribution. The intracellular fluorescence is due tobackground autofluorescence and not GFP.

[0067]FIG. 4B is a copy of a photographic representation showing thatthe K1544A mutant also lost polarized distribution of the fusionprotein, with the protein being detected in the apical and basolateralmembranes.

[0068]FIG. 4C is a copy of a photographic representation showing thatthe F1545A mutant has the same localization as the native protein.

[0069]FIG. 4D is a copy of a photographic representation showing thatthe triple mutant (i.e. T1543A K1544A F1545A) is localized apically inthe top down view (upper panel) however distributed in both the apicaland basolateral membranes in the XZ view (lower panel), indicating anon-polarized distribution.

[0070]FIG. 4E is a copy of a photographic representation showing thatthe S1542A mutant exhibits a less distinct distribution wherein theplasma membrane was outlined by the fluorescence of the protein, but oncloser inspection in the XZ view (lower panel), the fluorescence appearsto be in sub-membrane vesicles.

[0071]FIG. 5 is a copy of a photographic representation showing thedistribution of cMOAT polypeptides in L1210 cells, as follows:

[0072]FIG. 5A is a copy of a photographic representation showing L1210cells that were transiently transfected with cMOAT-gfp. The majority ofcMOAT-gfp accumulated in intracellular vesicles with minimal plasmamembrane localization.

[0073]FIG. 5B is a copy of a photographic representation showing L1210cells that were transiently transfected with ΔcMOAT-gfp. The majority ofΔcMOAT-gfp localized to the cell membrane.

[0074]FIG. 5C is a copy of a photographic representation showing M2 III6antibody binding to cMOAT in L1210 cells. Native cMOAT was detected inintracellular vesicles surrounding the nucleus (N). This localization isconsistent with cMOAT-gfp localization.

[0075]FIG. 5D is a copy of a photographic representation showing M2 III6antibody binding to ΔcMOAT in L1210 cells. ΔcMOAT was detected in thecell membrane confirming the effects of the TKF motif deletion foundwith ΔcMOAT-gfp.

[0076]FIG. 6 is a graphical representation showing the efflux of DNP-GSinto the supernatant by L1210 cells at specific time intervals, asdetermined by spectrophotometry (Olive et al (1994) Biochim. Biophys.Acta. 1224, 264-268). The control L1210 cells (o) and those cells thatwere transfected with wild type cMOAT (▪) had the same rate of efflux.The L1210 cells expressing ΔcMOAT (▴) had increased transport of theDNP-GS into the extracellular medium. The background transport of DNP-GSis due to constitutive MRP1. Results are the mean of three separateexperiments ± the S.D.

[0077]FIG. 7 is a graphical representation of an amino acid sequencealignment of the C terminal regions of ABC transporter proteins from anumber of species with the HisP protein. This alignment is derived froman alignment of the entire C-terminal cytoplasmic domain of 37 ABCtransporters. The cMOAT homologues have a distinct C-terminal extensionwhen compared with the basolaterally targeted proteins MRP1, MRP3, andMRP6. The sequences presented in the alignment are the C-terminalportions of the following naturally-occurring ABC transporter proteins:HisP (SEQ ID NO: 45); human cMOAT (SEQ ID NO: 17); mouse cMOAT (SEQ IDNO: 46); rat cMOAT (SEQ ID NO: 18); rabbit cMOAT (SEQ ID NO: 19); humanP-gp (SEQ ID NO: 20); rat P-gp (SEQ ID NO: 21); human MDR3 (SEQ ID NO:22); human MRP1 (SEQ ID NO: 23); and human MRP4 (SEQ ID NO: 47). The TKFmotif of each sequence is in bold type.

[0078]FIG. 8 is a copy of a photographic representation of a homologymodel of the C-terminal domains of native MRP1 (top panel) and nativecMOAT (lower panel), based on the crystal structure of HisP. The view islooking down on the subunit from the membrane into the cytoplasm. Thelower face is the C-terminal helix (marked “C-terminus” in each panel).The C-terminal helix of native cMOAT is clearly longer than in nativeMRP1. The T-K-F motif sits at the end of the C-terminal helix of cMOAT.

[0079]FIG. 9 is a graphical representation showing the enhancedresistance of L1210 cells expressing ΔcMOAT (i.e. SEQ ID NO: 4) to thechemotherapeutic agent Busulfan. Cells were incubated with a range ofconcentrations of Busulfan (x-axis) and the percentages of cellssurviving were determined, as indicated by the ordinate. Cells wereeither wild type cells (♦); L1210 cells expressing native cMOAT (-▪-);or L1210 cells expressing ΔcMOAT (-Δ-). The best-ft exponential curvefor L1210 cells expressing ΔcMOAT is also indicated. L1210 cellsexpressing ΔcMOAT had at least a 2-fold higher IC₅₀ for Busulfan thanthe other cells tested.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0080] One aspect of the present invention provides a modified ABCtransporter polypeptide having a novel distribution in the plasmamembrane of a cell compared to the corresponding native ABC transporterpolypeptide. It will be apparent from the preceding description that ABCtransporter polypeptides may have differential localization within theapical membranes of polarized and non-polarized cells. For example,native cMOAT, native MRP4, native P-gp, and native P-gp homologues (MDR2and MDR3) are generally found in the apical membrane domain of apolarized cell, such as hepatic cells. The native transporters thusfunction to transport organic anions across the canalicular membraneinto bile. The native polypeptides are also localized intracellularly innon-polarized cells. In contrast to these apical proteins, the MRP1,MRP3, and MRP6 polypeptides of humans are localized to the basolateralmembrane domain of polarized cells.

[0081] Accordingly, in a preferred embodiment, the present inventionencompasses modified forms of those ABC transporter polypeptides thatare normally found in the apical membrane of polarized cells. In aparticularly preferred embodiment, the modified ABC transporterpolypeptide of the invention is a modified cMOAT polypeptide, modifiedMDR3 polypeptide, or modified MRP4 polypeptide.

[0082] Preferably, the native ABC transporter polypeptide from which themodified ABC transporter polypeptide is derived is a polypeptide of ahuman or non-human mammal, such as, for example, a human, rat, rabbit,or mouse. In a particularly preferred embodiment, the polypeptide isfrom humans.

[0083] In accordance with this embodiment, it is particularly preferredthat the modified ABC transporter polypeptide of the invention consistof an amino acid sequence presented in any one of SEQ ID NOs: 4, 6, 10,12, 16, 48, or 49. A full description of each of said amino acidsequences is presented inter alia at pages 24 of the specification.Means for the production of these modified ABC transporter polypeptideswill be apparent from the exemplified subject matter described herein.

[0084] The modified ABC transporter polypeptide of the invention iscapable of accumulating in the plasma membrane of a polarized cell,however in contrast to the naturally-occurring form, the modified ABCtransporter polypeptide of the present invention is capable of beingdistributed predominantly to the basolateral membrane of a polarizedcell.

[0085] By “predominantly to the basolateral membrane” is meant that mostof said modified ABC transporter polypeptide is found in the basolateralmembrane of polarized cells. Preferably, more than about 70% of themodified ABC transporter is found in the basolateral membrane, and morepreferably, more than about 80%, and even more preferably, about 90% ofthe modified ABC transporter polypeptide is localized in the basolateralmembrane of polarized cells.

[0086] Polarized cell types will be well known to those skilled in theart. These include, for example, cultured epithelial cells such as MDCKcells, Caco-2 cells, and primary epithelial cells such as those cells ofhepatic and intestinal lineage, such as, for example, cells of thekidney, including the renal tubule; the liver; small intestine,including the small intestinal mucosa; liver; and pancreas.

[0087] In an alternative embodiment, the modified ABC transporterpolypeptide of the present invention accumulates in the plasma membraneof a non-polarized cell. The key observation by the inventors that themodified ABC transporter polypeptide of the invention accumulate in theplasma membrane of non-polarized cells is surprising and unexpected inview of the absence of detectable accumulation of thenaturally-occurring form in the plasma membranes of such cells.

[0088] Non-polarized cell types will be well known to those skilled inthe art. These include, for example, non-epithelial cells such as thoseforming the haematopoietic system and cultured cell types such as L1210cells and Jurkat cells.

[0089] Preferably, the modified ABC transporter polypeptide of thepresent invention consists of an active ABC transporter polypeptidecomprising a mutation wherein at least one amino acid residue in theC-terminal region of said active ABC transporter polypeptide issubstituted or deleted.

[0090] In the present context, the term “C-terminal region” or a similarterm, such as, for example, “C-terminus”, shall be taken to mean aportion comprising at least the C-terminal 20 amino acids of thecorresponding native or naturally-occurring ABC transporter polypeptide.Preferably a “C-terminal region” comprises at least the C-terminal 10amino acids of an ABC transporter polypeptide, and even more preferablyat least the C-terminal 5 amino acids of an ABC transporter polypeptide.

[0091] In a particularly preferred embodiment, a sequence comprisingthree amino acid residues in the C-terminal region of a naturallyoccurring ABC transporter polypeptide is mutated or deleted. As will beapparent from the subject matter described herein, a “C-terminal region”generally includes an amino acid sequence comprising a T-K-F-motif.

[0092] The term “T-K-F motif”, or similar term, shall be taken to referto an amino acid sequence derived from the amino acid sequence of an ABCtransporter polypeptide normally present in the apical membrane of apolarized cell, wherein said amino acid sequence is selected from thegroup consisting of:

[0093] (i) threonine-lysine-phenylalanine (i.e. T-K-F) (SEQ ID NO: 52);

[0094] (ii) threonine-alanine-phenylalanine (i.e. T-A-F)(SEQ ID NO: 53);

[0095] (iii) threonine-alanine-lysine (i.e. T-A-L) (SEQ ID NO: 54);

[0096] (iv) threonine-glutamate-leucine (i.e. T-E-L) (SEQ ID NO: 55);

[0097] (v) threonine-lysine-arginine (i.e. T-K-R) (SEQ ID NO: 56);

[0098] (vi) threonine-glutamine-asparagine (i.e. T-Q-N) (SEQ ID NO: 57);and

[0099] (vii) alanine-lysine-arginine (i.e. A-K-R) (SEQ ID NO: 58).

[0100] In this respect, the present inventors have demonstrated that aT-K-F motif as defined herein above is present in a number of ABCtransporter polypeptides that normally accumulate predominantly in theapical membrane of a polarized cell. It will also be understood that aT-K-F motif is not present in the C-terminal region of an ABCtransporter that normally accumulates predominantly in the basolateralmembrane of a polarized cell.

[0101] Surprisingly, mutation or deletion of the T-K-F motif of cMOAT,MDR3, or MRP4 alters the spatial accumulation of the modified ABCtransporter polypeptide within the plasma membrane of both polarized andnon-polarized cells. More particularly, mutation or deletion of theT-K-F motif produces a modified ABC transporter polypeptide capable ofaccumulating in the plasma membrane of a non-polarized cell orpredominantly in the basolateral membrane of a polarized cell. Thesemodified patterns of accumulation have utility in the field modifyingthe drug resistance of polarized and non-polarized cell types.

[0102] In an alternative embodiment, the modified ABC transporterpolypeptide of the present invention consists of an active ABCtransporter polypeptide comprising a mutation wherein at least one aminoacid residue of a tripeptide T-K-F motif present in said active ABCtransporter polypeptide is substituted or deleted. Preferably at leasttwo amino acid residues of the T-K-F motif is substituted or deleted.More preferably, all three amino acid residues of the T-K-F motif aredeleted or substituted. As will be apparent from the precedingdescription, such a substitution or deletion modifies the localizationof the modified ABC transporter polypeptide within the plasma membraneof both polarized and non-polarized cells.

[0103] The modified ABC transporter polypeptide may be a syntheticpeptide produced by any method known to those skilled in the art, suchas by using Fmoc chemistry. Alternatively, a modified ABC transporterpolypeptide may be produced by recombinant means, wherein nucleic acidencoding a native ABC transporter polypeptide is subjected tomutagenesis and the mutated sequence is expressed in a cell to producethe modified ABC transporter polypeptide.

[0104] Substitutions encompass any amino acid alterations in which anamino acid is replaced with a different conventional or non-conventionalamino acid residue. To produce the modified ABC transporter polypeptideof the invention, amino acids in the C-terminal region of a native ABCtransporter polypeptide may be substituted for other conventional ornon-conventional amino acids having different properties. For examplethe new amino acid may have a different property to the base amino acidthat is selected from the group consisting of hydrophobicity,hydrophilicity, hydrophobic moment, antigenicity, and propensity to formor break α-helical structures or β-sheet structures.

[0105] Conventional amino acid residues contemplated herein aredescribed in Table 1. Non-conventional amino acid residues contemplatedherein are described in Table 2.

[0106] Substitutions encompassed by the present invention will generallybe “non-conservative”. This means that an amino acid residue which ispresent in a native ABC transporter polypeptide is substituted with anamino acid having a different property. Such non-conservativesubstitutions generally involve a substitution for an amino acid from adifferent group to the base amino acid. For example a non-chargedresidue can be substituted for a charged residue, or a hydrophobicresidue can be substituted for alanine.

[0107] Particularly preferred amino add substitutions are selected fromthe group consisting of Ser

Ala; Thr

Ala; Lys

Ala or Pro; and Phe

Ala or Val.

[0108] Amino acid substitutions may be of multiple residues, eitherclustered or dispersed, within the C-terminal region, and preferably arepositioned within the T-K-F motif of the native ABC transporterpolypeptide or immediately adjacent thereto. Accordingly, the clusteredsubstitution of Thr-Lys-Phe (i.e. the T-K-F motif) for Ala-Ala-Ala isclearly within the scope of this invention.

[0109] Amino acid deletions are those mutations wherein one or moreamino acid residues within the C-terminal region of an ABC transporterpolypeptide including the T-K-F motif, are removed. Amino acid deletionswill usually be of the order of about 1-10 amino acid residues.

[0110] Amino acid insertions are those mutations wherein one or moreamino acid residues are added to C-terminal region of an ABC transporterpolypeptide, preferably disrupting the T-K-F motif. TABLE 1 Three-letterOne-letter Amino Acid Abbreviation Symbol Alanine Ala A Arginine Arg RAsparagine Asn N Aspartic acid Asp D Cysteine Cys C Glutamine Gln QGlutamic acid Glu E Glycine Gly G Histidine His H Isoleucine Ile ILeucine Leu L Lysine Lys K Methionine Met M Phenylalanine Phe F ProlinePro P Serine Ser S Threonine Thr T Tryptophan Trp W Tyrosine Tyr YValine Val V Any amino acid as above Xaa X

[0111] TABLE 2 Non-conventional Non-conventional amino acid Code aminoacid Code α-aminobutyric acid Abu L-N-methylalanine Nmalaα-amino-α-methylbutyrate Mgabu L-N-methylarginine Nmargaminocyclopropane- Cpro L-N-methylasparagine Nmasn carboxylateL-N-methylaspartic acid Nmasp aminoisobutyric acid AibL-N-methylcysteine Nmcys aminonorbornyl- Norb L-N-methylglutamine Nmglncarboxylate L-N-methylglutamic acid Nmglu cyclohexylalanine ChexaL-N-methylhistidine Nmhis cyclopentylalanine Cpen L-N-methylisolleucineNmile D-alanine Dal L-N-methylleucine Nmleu D-arginine DargL-N-methyllysine Nmlys D-aspartic acid Dasp L-N-methylmethionine NmmetD-cysteine Dcys L-N-methylnorleucine Nmnle D-glutamine DglnL-N-methylnorvaline Nmnva D-glutamic acid Dglu L-N-methylornithine NmornD-histidine Dhis L-N-methylphenylalanine Nmphe D-isoleucine DileL-N-methylproline Nmpro D-leucine Dleu L-N-methylserine Nmser D-lysineDlys L-N-methylthreonine Nmthr D-methionine Dmet L-N-methyltryptophanNmtrp D-ornithine Dorn L-N-methyltyrosine Nmtyr D-phenylalanine DpheL-N-methylvaline Nmval D-proline Dpro L-N-methylethylglycine NmetgD-serine Dser L-N-methyl-t-butylglycine Nmtbug D-threonine DthrL-norleucine Nle D-tryptophan Dtrp L-norvaline Nva D-tyrosine Dtyrα-methyl-aminoisobutyrate Maib D-valine Dval α-methyl-γ-aminobutyrateMgabu D-α-methylalanine Dmala α-methylcyclohexylalanine MchexaD-α-methylarginine Dmarg α-methylcylcopentylalanine McpenD-α-methylasparagine Dmasn α-methyl-α-napthylalanine ManapD-α-methylaspartate Dmasp α-methylpenicillamine Mpen D-α-methylcysteineDmcys N-(4-aminobutyl)glycine Nglu D-α-methylglutamine DmglnN-(2-aminoethyl)glycine Naeg D-α-methylhistidine DmhisN-(3-aminopropyl)glycine Norn D-α-methylisoleucine DmileN-amino-α-methylbutyrate Nmaabu D-α-methylleucine Dmleu α-napthylalanineAnap D-α-methyllysine Dmlys N-benzylglycine Nphe D-α-methylmethionineDmmet N-(2-carbamylethyl)glycine Ngln D-α-methylornithine DmornN-(carbamylmethyl)glycine Nasn D-α-methylphenylalanine DmpheN-(2-carboxyethyl)glycine Nglu D-α-methylproline DmproN-(carboxymethyl)glycine Nasp D-α-methylserine Dmser N-cyclobutylglycineNcbut D-α-methylthreonine Dmthr N-cycloheptylglycine NchepD-α-methyltryptophan Dmtrp N-cyclohexylglycine Nchex D-α-methyltyrosineDmty N-cyclodecylglycine Ncdec D-α-methylvaline DmvalN-cylcododecylglycine Ncdod D-N-methylalanine Dnmala N-cyclooctylglycineNcoct D-N-methylarginine Dnmarg N-cyclopropylglycine NcproD-N-methylasparagine Dnmasn N-cycloundecylglycine NcundD-N-methylaspartate Dnmasp N-(2,2-diphenylethyl)glycine NbhmD-N-methylcysteine Dnmcys N-(3,3-diphenylpropyl)glycine NbheD-N-methylglutamine Dnmgln N-(3-guanidinopropyl)glycine NargD-N-methylglutamate Dnmglu N-(1-hydroxyethyl)glycine NthrD-N-methylhistidine Dnmhis N-(hydroxyethyl))glycine NserD-N-methylisoleucine Dnmile N-(imidazolylethyl))glycine NhisD-N-methylleucine Dnmleu N-(3-indolylyethyl)glycine NhtrpD-N-methyllysine Dnmlys N-methyl-γ-aminobutyrate NmgabuN-methylcyclohexylalanine Nmchexa D-N-methylmethionine DnmmetD-N-methylornithine Dnmorn N-methylcyclopentylalanine NmcpenN-methylglycine Nala D-N-methylphenylalanine DnmpheN-methylaminoisobutyrate Nmaib D-N-methylproline DnmproN-(1-methylpropyl)glycine Nile D-N-methylserine DnmserN-(2-methylpropyl)glycine Nleu D-N-methylthreonine DnmthrD-N-methyltryptophan Dnmtrp N-(1-methylethyl)glycine NvalD-N-methyltyrosine Dnmtyr N-methyla-napthylalanine NmanapD-N-methylvaline Dnmval N-methylpenicillamine Nmpen γ-aminobutyric acidGabu N-(p-hydroxyphenyl)glycine Nhtyr L-t-butylglycine TbugN-(thiomethyl)glycine Ncys L-ethylglycine Etg penicillamine PenL-homophenylalanine Hphe L-α-methylalanine Mala L-α-methylarginine MargL-α-methylasparagine Masn L-α-methylaspartate MaspL-α-methyl-t-butylglycine Mtbug L-α-methylcysteine McysL-methylethylglycine Metg L-α-methylglutamine Mgln L-α-methylglutamateMglu L-α-methylhistidine Mhis L-α-methylhomophenylalanine MhpheL-α-methylisoleucine Mile N-(2-methylthioethyl)glycine NmetL-α-methylleucine Mleu L-α-methyllysine Mlys L-α-methylmethionine MmetL-α-methylnorleucine Mnle L-α-methylnorvaline Mnva L-α-methylornithineMorn L-α-methylphenylalanine Mphe L-α-methylproline MproL-α-methylserine Mser L-α-methylthreonine Mthr L-α-methyltryptophan MtrpL-α-methyltyrosine Mtyr L-α-methylvaline MvalL-N-methylhomophenylalanine Nmhphe N-(N-(2,2-diphenylethyl) NnbhmN-(N-(3,3-diphenylpropyl) Nnbhe carbamylmethyl)glycinecarbamylmethyl)glycine

[0112] In a particularly preferred embodiment of the invention, 14 aminoacid residues is deleted from the C-terminus of a cMOAT polypeptide,P-gp polypeptide, MDR3 polypeptide, or MRP4 polypeptide, to produce amodified ABC transporter polypeptide. Alternatively, at least the firstof second amino acid residue of the presumptive T-K-F motif is deletedor substituted. As exemplified herein for cMOAT, mutation or deletion ofT1543 and/or K1544, optionally further including a mutation or deletionof F1545, significantly modifies protein targeting. Also exemplifiedherein, deletion of the entire T-K-F motif of cMOAT, MDR3, or MRP4modified cellular localization of the protein.

[0113] Accordingly, a particularly preferred embodiment of the inventionprovides a modified ABC transporter polypeptide consisting of a modifiedcMOAT polypeptide having an amino acid sequence substantially as setforth in any one of SEQ ID NOs: 4, 6, 10, 12, 16, 48, or 49, or afunctional variant thereof having up to 5 amino adds removed from theC-terminal region and preferably, having as many as 10-20 amino acidsremoved from the C-terminal region of the corresponding native protein.In the present context, the term “functional variant” means any modifiedABC transporter polypeptide that has the transport function of a nativeABC transporter polypeptide notwithstanding that it is localized in adifferent membrane domain to the native ABC transporter polypeptide.

[0114] This aspect of the invention clearly includes any fusion proteincomprising the modified ABC transporter, particularly a fusionpolypeptide between the modified ABC transporter and green fluorescentprotein (GFP) as exemplified herein.

[0115] A second aspect of the invention clearly extends to the isolatednucleic acid encoding the modified ABC transporter polypeptide describedherein.

[0116] This aspect of the invention relates to a nucleic acid moleculeconsisting of a nucleotide sequence encoding a functional ABCtransporter polypeptide, wherein a native ABC transporterpolypeptide-encoding nucleotide sequence has a mutation selected fromthe group consisting of:

[0117] (i) a deletion of at least nine nucleotides from the 3′-end ofthe coding region of the wild-type gene sequence;

[0118] (ii) a deletion from the 3′-end of the coding region of thewild-type gene sequence which removes at least a part of the nucleotidesequence of said gene encoding the T-K-F motif;

[0119] (iii) a substitution within the 3′-end of the coding region ofthe wild-type gene which mutates the nucleotide sequence of said geneencoding the T-K-F motif;

[0120] (iv) an insertion within the 3′-end of the coding region of thewild-type gene which mutates the nucleotide sequence of said geneencoding the T-K-F motif; and

[0121] (v) any mutation that introduces a stop codon within the 3-end ofthe coding region of the wild-type gene thereby preventing nucleotidesequences encoding TKF motif of said gene from being translated.

[0122] Preferably, the deletion referred to in sub-paragraph (i) supracomprises a deletion of at least about 10 nucleotides, more preferably,at least about 11 nucleotides, and more preferably at least about 12nucleotides from the 3′-end of the coding region of the correspondingnative ABC transporter polypeptide-encoding nucleotide sequence.

[0123] In a particularly preferred embodiment, the isolated nucleic acidof the invention consists of the nucleotide sequence of the modifiedcMOAT-encoding gene set forth in any one of SEQ ID NOs: 3, 5, 9, 11, or15.

[0124] In an alternative embodiment, nucleic acid encoding a modifiedABC transporter polypeptide is produced by amplification using primerscontaining mutations therein, as described in the examples. As will beknown to those skilled in the art, the amplified mutant sequence willinclude the nucleotide sequence of the primer, or the complementarysequence thereto at the 3′-end of its coding region. Accordingly, thepresent invention clearly encompasses a modified ABC transporter thatincludes a nucleotide sequence selected from the group consisting of SEQID Nos: 26 to 33, 37, 59-62, and a complementary nucleotide sequence toany one of said SEQ ID NOs.

[0125] To express the modified ABC transporter polypeptide of thepresent invention in a cell, such as a mammalian cell, it is desirableto place the nucleic acid molecule in an expressible format in operableconnection with a suitable promoter sequence.

[0126] As used herein, a “nucleic acid molecule in an expressibleformat” comprises the protein-encoding region in operable connectionwith a promoter or other regulatory sequence capable of regulatingexpression of the modified ABC transporter polypeptide encoded by saidprotein-encoding region. As will be known tot hose skilled in the art,such expression is generally carried out in an appropriate cell host.

[0127] Reference herein to a “promoter” is to be taken in its broadestcontext to include the transcriptional regulatory sequences of aclassical genomic gene. Such regulatory sequences include the TATA boxwhich is required for accurate transcription initiation, with or withouta CCAAT box sequence and additional regulatory elements (i.e., upstreamactivating sequences, enhancers and silencers) that alter geneexpression in response to developmental and/or external stimuli, or in atissue-specific manner. In the present context, the term “promoter” isalso used to describe a recombinant, synthetic or fusion molecule, orderivative that is capable of conferring, activating or enhancingexpression of nucleic acid encoding the modified ABC transporterpolypeptide of the invention. Preferred promoters can contain additionalcopies of one or more specific regulatory elements to further enhanceexpression and/or to alter the spatial expression and/or temporalexpression of the said nucleic acid molecule.

[0128] Placing a nucleic acid molecule under the regulatory control of(i.e., “in operable connection with”) a promoter sequence meanspositioning the said molecule such that expression is controlled by thepromoter sequence. Promoters are generally, but not necessarily,positioned 5′ (upstream) to the genes that they control. To produce aheterologous promoter/structural gene combination, the promoter isgenerally positioned at a distance from the gene transcription startsite that is approximately the same as the distance between thatpromoter and the gene it controls in its natural setting. Furthermore,the regulatory elements comprising a promoter are usually positionedwithin 2 kb of the start site of transcription of the gene. As is knownin the art, some variation in this distance can be accommodated withoutloss of promoter function. Similarly, the preferred positioning of aregulatory sequence element with respect to a heterologous gene to beplaced under its control is defined by the positioning of the element inits natural setting, i.e., the genes from which it is derived. Again, asis known in the art, some variation in this distance can also occur.

[0129] Preferably, the promoter sequence facilitates expression of themodified ABC transporter polypeptide in a bacterial cell, yeast, fungalcell, insect cell, or mammalian cell.

[0130] The prerequisite for producing intact polypeptides in bacteriasuch as E. coli is the use of a strong promoter with an effectiveribosome binding site. Typical promoters suitable for expression inbacterial cells such as E. coli include, but are not limited to, thelacz promoter, temperature-sensitive λ_(L) or λ_(R) promoters, T7promoter or the IPTG-inducible tac promoter. A number of other vectorsystems for expressing the nucleic acid molecule of the invention in E.coli are well-known in the art and are described, for example, inAusubel et al (1987). In: Current Protocols in Molecular Biology. WileyInterscience (ISBN 047150338) or Sambrook et al (1989) MolecularCloning: A Laboratory Manual, Cold Spring Harbor Laboratory, Cold SpringHarbor, N.Y. Numerous plasmids with suitable promoter sequences forexpression in bacteria and efficient ribosome binding sites have beendescribed, such as for example, pKC30 (λ_(L): Shimatake and Rosenberg(1981) Nature 292, 128), pKK173-3 (tac: Amann and Brosius (1985). Gene40,183), pET-3 (T7: Studier and Moffat (1986) J. Mol. Biol. 189,113),the pFLEX series of expression vectors (Pfizer Inc., CT, USA) or the pQEseries of expression vectors (Qiagen, Calif.), amongst others.

[0131] Suitable promoters for use in eukaryotic expression vectorsinclude those capable of regulating expression in mammalian cells,insect cells such as Sf9 or Sf21 (Spodoptera furgiperda) cells, yeastcells and fungal cells. Preferred promoters for expression in eukaryoticcells include the p10 promoter, MMTV promoter, polyhedron promoter, theSV40 early promoter and the cytomegalovirus (CMV-IE) promoter, promotersderived from immunoglobulin-producing cells (see, U.S. Pat. No.4,663,281), polyoma virus promoters, and the LTR from variousretroviruses (such as murine leukemia virus, murine or Rous sarcomavirus and HIV), amongst others (See, Enhancers and Eukaryotic GeneExpression, Cold Spring Harbor Press, New York, 1983, which isincorporated herein by reference). Examples of other expression controlsequences are enhancers or promoters derived from viruses, such as SV40,Adenovirus, Bovine Papilloma Virus, and the like.

[0132] A preferred expressible format for the modified ABC transporterpolypeptide of the invention is achieved by placing the nucleotidesequence encoding said polypeptide and a promoter to which it isoperably connected within a gene expression construct or vector.

[0133] Accordingly, a further aspect of the present invention provides agene construct comprising a nucleotide sequence encoding the modifiedABC transporter polypeptide of the invention.

[0134] The gene construct is preferably a plasmid or a retrovirusvector. Numerous expression vectors suitable for the present purposehave been described and are readily available. The expression vector maybe based upon the pcDNA3 vector (Medos Company Pty Ltd, Victoria,Australia) that comprises the CMV promoter and BGH terminator sequences.Alternatively, the SG5 expression vector (Greene et al. (1988) NucleicAcids Res. 15, 369; Stratagene), or the pQE series of vectors (Qiagen)are particularly useful for such purposes.

[0135] A preferred mammalian plasmid-based gene expression construct isthe pRc/CMV plasmid (Invitrogen), which utilizes the CMV promoter todrive expression in mammalian host cells. Alternatively, a retroviralexpression vector containing the Harvey murine sarcoma virus (Ha-MSV)long terminal repeats (LTRs) flanking the promoter and nucleic acidencoding the modified ABC transporter polypeptide may be used. Onepreferred Ha-MSV is the pC01 expression vector.

[0136] The gene constructs described herein may further comprise geneticsequences corresponding to a bacterial origin of replication and/or aselectable marker gene suitable for the maintenance and replication ofsaid gene construct in a prokaryotic or eukaryotic cell, tissue ororganism. Such sequences are well known in the art.

[0137] Selectable marker genes include genes which when expressed arecapable of conferring resistance on a cell to a compound which would,absent expression of said selectable marker gene, prevent or slow cellproliferation or result in cell death. Those skilled in the art areaware that various antibiotic-resistance genes, such as those conferringresistance to ampicillin, Claforan, gentamycin, G418, hygromycin,rifampicin, kanamycin, neomycin, spectinomycin, or tetracycline, aregenerally used in such gene constructs as selectable markers.

[0138] The origin of replication and/or a selectable marker gene ispreferably separated from the coding sequences that encode the modifiedABC transporter polypeptide.

[0139] Methods for the production of recombinant plasmids, cosmids,bacteriophage molecules or other recombinant molecules are well known tothose of ordinary skill in the art and can be accomplished without undueexperimentation.

[0140] The gene constructs of the invention, including any expressionvectors, are capable of introduction into, and expression in, an invitro cell culture, or for introduction into, with or withoutintegration into the genome of a cultured cell, cell line and/ortransgenic animal.

[0141] In a particularly preferred embodiment contemplated herein, thegene constructs are used in gene therapy to transfer nucleic acidencoding the modified ABC transporter polypeptide to human cells.Preferably, such transfer is for the purposes of transplanting humancells expressing the modified ABC transporter polypeptide to humansduring somatic therapy. Gene delivery systems may be viral, such as, forexample, using retrovirus-based vectors or Adenovirus-based, oralternatively, a non-viral delivery system may be used, including anyplasmid DNA-based delivery systems. For example, human haemopoieticcells or bone marrow cells or cells of the gastrointestinal tract aretransfected with Ad21 or other adenovirus expressing the modified ABCtransporter of the invention, and the transfected cells transplantedinto the appropriate organ of a human patient to enhance drug resistancein that organ. Methods for performing somatic gene therapy are known tothose skilled in the art (Fibison (2000) Nurs. Clin. North Am. 35,757-772).

[0142] The present invention also provides a transformed cell comprisingthe nucleic acid molecule of the invention.

[0143] As used herein, unless the context requires otherwise, the word“cell” shall be taken to include a clonal or non-clonal group of cells.A group of cells may be functionally organized into whole tissue, anorgan, or organism, or into a part of said tissue, organ or organism.The term “cell” shall further include any cell lysate of an isolatedcell or group of cells.

[0144] As used herein, the term “transformed cell” is meant to alsoinclude the progeny of a transformed cell.

[0145] The host cell may be a mammalian cell, more preferably a humancell, canine cell, rat cell, rabbit cell or murine cell, and even morepreferably the cell is a drug-sensitive primary epithelial cell ornon-epithelial cell of humans, such as, for example, a bone marrow cell,a cell of the gastrointestinal tract, or a cell of the haematopoieticsystem.

[0146] Examples of eukaryotic cell lines contemplated herein to beuseful include NIH 3T3, COS, VERO, HeLa, mouse C127, mouse L1210,Chinese hamster ovary (CHO), WI-38, baby hamster kidney (BHK), and MDCKcell lines. Such cell lines are readily available to those skilled inthe art.

[0147] In a particularly preferred embodiment, the host cell is anon-polarized, such as, for example, the murine leukaemia cell lineL1210, or alternatively, a polarized cell, such as an MDCK cell.

[0148] Means for introducing the isolated nucleic acid molecule or agenetic construct comprising same into a cell for expression of theimmunogenic component of the vaccine composition are well known to thoseskilled in the art. The technique used for a given organism depends onthe known successful techniques. Means for introducing recombinant DNAinto animal cells include microinjection, transfection mediated byDEAE-dextran, transfection mediated by liposomes such as by usinglipofectamine (Gibco, Md., USA) and/or cellfectin (Gibco, Md., USA),PEG-mediated DNA uptake, electroporation and micropartide bombardmentsuch as by using DNA-coated tungsten or gold particles (Agracetus Inc.,WI, USA).

[0149] Moreover, transfection of a mammalian cell with the geneconstruct of the present invention results in the transformation ofpolarized and non-polarized cells from a drug-sensitive phenotype to adrug-resistant phenotype. Thus, cells that would normally be damaged orkilled by certain drugs, such as chemotherapeutic agents (e.g.busulfan), are able to tolerate much higher levels of drug exposure withlittle or no adverse effect The cells thus acquire a multidrug resistantphenotype comparable to that observed in tumor cells subjected tovarious chemotherapeutic agents.

[0150] Accordingly, the gene construct according to this embodiment ofthe invention is particularly useful for conferring novel drugresistance characteristics on a cell, in particular a non-polarizedcell, or alternatively, for transporting particular drugs from the cell.

[0151] Wherein the cell is a non-polarized cell, such as, for example,certain non-epithelial cells including fibroblasts and cells of thehaemopoietic system, the modified ABC transporter polypeptide islocalized generally within the plasma membrane. This confers resistanceon the non-polarized cell, which would otherwise have a reduced effluxcapacity.

[0152] Alternatively, wherein the cell is a polarized cell, such as, forexample, a cultured epithelial cell (e.g. MDCK, Caco-2) or a primaryepithelial cell (e.g. hepatocytes, intestinal cell, hippocampalneurons), the modified ABC transporter polypeptide is surprisinglydistributed predominantly to the basolateral membrane. Localization ofthe modified ABC transporter polypeptide to the basolateral membranes ofa polarized cell facilitates the efflux of certain ligand drugs from thecell via the basolateral membrane to confer resistance propertiesthereon.

[0153] It is highly desirable for the expressed modified ABC transporterpolypeptide to confer resistance on the cell to one or more chemicalcompounds, such as, for example, a cytostatic or cytotoxic compound usedin the treatment of infection or disease. For example, protection ofnon-polarized cells (e.g. cells of the haematopoietic system) isdesirable during the treatment of patients with cytotoxic or cytostaticcompounds.

[0154] As used herein, the term “chemical compound” shall be taken tomean any natural product, or synthetic compound having a definablechemical structure, and, in particular, a natural product or syntheticcompound that is capable of being actively-transported into or out of acell. Those skilled in the art will be aware that active-transportrefers to an energy-dependent transport process, such as, for example, atransport process utilizing ATP or GTP or a nucleoside analogue thereof.Preferably, the chemical compounds against which resistance orsensitivity is modulated in accordance with the invention are thosechemical compounds that are transported via ABC transporters, membranetransporters, or like transporters.

[0155] More preferably, the chemical compounds against which resistanceor sensitivity is modulated in accordance with the invention are naturalproducts or synthetic compounds. These are also useful in the treatmentand/or prophylaxis of a disease of humans or other animals, such as, forexample, anti-bacterial, anti-fungal, and, more preferably,chemotherapeutic agents.

[0156] Preferred anti-bacterial agents are antibiotic compounds.Antibiotics include quinolone antibiotics, sulfonamide antibiotics,cephalosporin antibiotics, or aminoglycoside antibiotics. these may beselected from the group consisting of acyclovir, adriamycin, antimycin,amikacin, amoxicillin, amoxicillin/clavulanate (augmentin), amphotericinb (fungizone), ampicillin, atovaquone (mepron), azithromycin(zithromax), cefazolin, cefepime (maxipime), ceftazidime, cefotaxime(claforan), cefotetan (cefotan), cefpodoxime (vantin), ceftizoxime(cefizox), ceftriaxone (rocephin), cefuroxime (zinacef), cephalexin,clotrimazole (mycelex), ciprofloxacin (cipro), clarithromycin (biaxin),clindamycin (cleocin), oxycycline, erythromycin lactobionate,famciclovir (famvir), fluconazole (diflucan), foscamet (foscavir),ganciclovir, gentamicin, imipenem/cilastatin (primaxin), isoniazid,itraconazole (sporanox), nafcillin, nitrofurantoin, nystatin,oligomycin, paromomycin, penicillin g, piperacillin/tazobactam (zosyn),rifampin (rifadin), ticarcillin/clavulanate (timentin), tobramycin,trimethoprim sulfamethoxazole, valacyclovir (valtrex), and vancomycin.The invention also extends to conferring resistance against the chloridesalts or sulfated derivatives of the antibiotics supra, or against anyderivative or related compound.

[0157] Preferred anti-fungal compounds are imidazoles (includingbifonazole [i.e. 1-(α-biphenyl-4-ylbenzyl)imidazole], clotrimazole,intraconazole, fluconazole, econazole nitrate, ketoconazole, astemizole,metronidazole (flagyl) and miconazole nitrate [i.e.1-[2,4-dichloro-β-(2,4-dichlorobenzyloxy)phenethyl]imidazole nitrate]);allylamines (including terbafine hydrochloride and terfenadine);amorolfine hydrochloride(cis-4-[(RS)3-[4(1,1-dimethylpropyl)phenyl]-2-methylpropyl]-2,6-dimethyl morpholine hydrochloride) and chloride salts andsulfated derivatives thereof, and derivatives and related compoundsthereto.

[0158] A “chemotherapeutic agent” is a cytostatic and/or cytotoxiccompound that is capable of rendering a mammalian cell inviable (i.e. acytotoxin). A chemotherapeutic agent will at least reduce the capacityof a cell to grow and/or to proliferate (i.e. a cytostat). The cytotoxicor cytostatic properties of chemotherapeutic agents confer utility onthese compounds in the therapeutic or prophylactic treatment of acancerous or pre-cancerous cell, or a tumor, in an animal.

[0159] Preferred chemotherapeutic agents are selected from the groupconsisting of: busulphan (busulfan), cisplatin, cyclophosphamide,chlorambucil, BCNU, melphalan, merchlorethane, vinblastine sulphate, andetoposide (VP-16. VP-16-213, or VePesid). Other chemotherapeutic agentsinclude vinca alkaloids selected from the group consisting of:vincristine sulfate, oncovin, velban, velsar, taxol, andepipodophyllotoxin (including podophyllotoxin and the syntheticderivatives thereof, teniposide (VM-26). The estrogen receptorantagonist tamoxifen, and the anti-neoplastic antibiotics adriamycin,bleomycin, doxorubicin, daunorubicin, daunomycin, rubidomycin,cerubidine, daunoblastina, plicomycin, and mitoxanthrone, and chloridesalts and sulfated derivatives thereof, and related compounds thereto,are also useful in chemotherapy.

[0160] A further aspect of the invention provides a method of enhancingthe resistance of a cell to a chemical compound comprising expressing amodified ABC transporter polypeptide in said cell for a time and underconditions sufficient for said cell to have modified growth and/orviability in the presence of said compound. Cell viability assays havebeen described in detail (Cui et al (1999) Mol. Pharmacol. 55, 929-937)and are readily adapatable to determining the enhanced resistance ofcells expressing the modified ABC transporters of the invention.

[0161] This embodiment of the present invention clearly encompasses theconferring of enhanced growth and/or viability in the presence of thechemical compound or drug being tested. By virtue of its retainedability to enhance efflux of any substrate, in combination with itsdifferent membrane localization compared to the corresponding native ABCtransporter, the modified ABC transporter of the invention enhancesefflux of cytotoxic/cytostatic compounds compared to the correspondingnative ABC transporter. Whilst not being bound by any theory or mode ofaction, the compound may be conjugated to glutathione, glucuronate, orsulfate, before it is transported from the cell.

[0162] For example, efflux of a cytotoxic/cytostatic drug substrate froma transfected polarized cell that expresses both the modified ABCtransporter and the corresponding endogenous native ABC transporter willoccur via both the apical and basolateral membranes, thereby enhancingtotal efflux compared to a non-transfected polarized cell.

[0163] Similarly, efflux of a cytotoxic/cytostatic drug substrate from atransfected non-polarized cell that expresses both the modified ABCtransporter and the corresponding endogenous native ABC transporter willoccur via the plasma membrane rather than being localized in theintracellular vesicles, thereby enhancing total efflux compared to anon-transfected non-polarized cell. As exemplified herein, ectopicexpression of the modified cMOAT polypeptide of the invention enhancesresistance of L1210 cells to Busulfan compared to non-transected L1210cells.

[0164] The distribution pattern of naturally-occurring ABC transporterpolypeptides in the tissues of humans or mammals provides for theextension of this aspect of the invention to further include thesite-specific enhancement of drug resistance in humans and animals.According to this embodiment of the invention, the modified ABCtransporter polypeptide of the invention is used in combination with oneor more inhibitors of an ABC transporter which is different to that fromwhich said modified ABC transporter polypeptide is derived (i.e. aheterologous ABC transporter polypeptide).

[0165] Accordingly, a further aspect of the invention provides a methodof protecting a non-polarized cell of an organism or tissue comprisingsaid non-polarized cell during the administration of a cytotoxic orcytostatic chemical compound to a subject, said method comprising:

[0166] (i) expressing a modified ABC transporter polypeptide in saidnon-polarized cell for a time and under conditions sufficient for saidcell to efficiently transport said cytotoxic or cytostatic compound fromsaid cell or otherwise acquire resistance to said compound; and

[0167] (ii) optionally, administering one or more inhibitors of an ABCtransporter for a time and under conditions sufficient for ablating orinhibiting the growth of the cell expressing said ABC transporter,wherein said ABC transporter is different to that from which saidmodified ABC transporter polypeptide is derived (i.e. a heterologous ABCtransporter polypeptide) and is involved in the transport of saidcytotoxic or cytostatic chemical compound.

[0168] Preferably, the cell of sub-paragraph (ii) supra is a polarizedcell or a non-polarized tumor cell. Preferably, the non-polarized cellof a sub-paragraph (i) supra is a cell of the haematopoietic system.

[0169] In a particularly preferred embodiment, the cytotoxic/cytostaticcompound is a chemotherapeutic agent, such as, for example, Busulfan.

[0170] For example, modified cMOAT can be used to protect thehaematopoietic system during chemotherapy that ablates non-haemopoietictumor cells. Preferably during such therapeutic regimes, one or moreP-gp antagonists can also be administered to inhibit P-gp activity innon-haemopoietic cells, to enhance the efficacy of the chemotherapeuticagent. Wherein P-gp activity is also inhibited, it is particularlypreferred that such inhibition is in respect of endogenous P-gp activityin an epithelial tumor cell or alternatively, in a non-polarized tumorcell that over-expresses P-gp.

[0171] Similarly, a modified cMOAT polypeptide can be used to protectthe haematopoietic system, preferably in conjunction with one or moreMDR antagonists to inhibit MDR activity in the apical membrane of anon-hematological tumor cell, and one or more chemotherapeutic agents toinhibit tumorigenesis.

[0172] Similarly, a modified cMOAT polypeptide can be used to protectthe haematopoietic system, preferably in conjunction with one or moreantagonists to inhibit the activity of MRP1 and its homologues in thebasolateral membrane of tumor cells, and one or more chemotherapeuticagents to ablate tumor cells.

[0173] In an alternative embodiment, a modified MDR3 polypeptide can beused to protect the haematopoietic system, preferably in conjunctionwith one or more cMOAT antagonists to inhibit cMOAT activity and/or oneor more antagonists to inhibit MDR homologue activity in the membrane ofnon-hematological tumor cells and/or one or more antagonists to inhibitthe activity of MRP1 and its homologues in the basolateral membrane oftumor cells, and one or more chemotherapeutic agents to ablate tumorcells.

[0174] In an alternative embodiment, a modified MDR homologuepolypeptide can be used to protect the haematopoietic system, preferablyin conjunction with one or more cMOAT antagonists to inhibit cMOATactivity and/or one or more P-gp antagonists to inhibit P-gp activity inthe membrane of non-hematological tumor cells and/or one or moreantagonists to inhibit the activity of MRP1 and its homologues in thebasolateral membrane of tumor cells, and one or more chemotherapeuticagents to ablate tumor cells.

[0175] It will be apparent from the preceding description that amodified cMOAT polypeptide or modified MDR3 polypeptide or modified MRP4polypeptide can be used to confer resistance in any non-polarized cellin which the corresponding naturally-occurring ABC transporterpolypeptide is not present or active. In such embodiments it will alsobe apparent that the invention does not require simultaneous orconsequential inhibition of endogenous ABC transporter activity innon-hematological tumor cells, notwithstanding that this feature isclearly encompassed by the invention.

[0176] The present invention further provides for the enhancement ofdrug resistance in a polarized cell in which the correspondingnaturally-occurring ABC transporter polypeptide is already present oractive in the apical membrane domain, preferably alongside the use ofone or more ABC transporter antagonists to inhibit a heterologous ABCtransporter polypeptide activity in tumorigenic non-polarized cells, andthe use of one or more chemotherapeutic agents to ablate the tumor.

[0177] Accordingly, in an alternative embodiment, the present inventionprovides a method of enhancing the resistance of a polarized cell of anorganism or tissue comprising said polarized cell during theadministration of a cytotoxic or cytostatic chemical compound to asubject, said method comprising:

[0178] (i) introducing or expressing a modified ABC transporterpolypeptide in said polarized cell for a time and under conditionssufficient for said cell to enhance transport said cytotoxic orcytostatic compound from said cell or otherwise enhance resistance tosaid compound; and

[0179] (ii) optionally, administering one or more inhibitors of an ABCtransporter of a cell for a time and under conditions sufficient forablating or inhibiting the growth of said cell, wherein said ABCtransporter is different to that from which said modified ABCtransporter polypeptide is derived (i.e. a heterologous ABC transporterpolypeptide) and is involved in the transport of said cytotoxic orcytostatic chemical compound.

[0180] Preferably, the cell of subparagraph (ii) supra is anon-polarized cell. Preferably, the polarized cell is a primaryepithelial cell (e.g. hepatocyte, intestinal cell, or hippocampalneuron, amongst others).

[0181] Preferred inhibitors of the cMOAT polypeptide and homologouspolypeptides of other species are listed in Table 3. TABLE 3 INHIBITORSOF cMOAT POLYPEPTIDES Cholestatic agents: α-Naphthylisothiocyanate,Chlorpromazine, Cyclosporin, Estradiol-17β- glucuronide,Ethinylestradiol, Glycolithocholate-3α-O-sulfate,Lithocholate-3α-O-glucuronide, Manganese-bilirubin, Phalloidin,Taurocholate, Taurolithocholate

[0182] The present invention clearly contemplates the administration ofa cytostatic compound or cytotoxic compound to a subject, wherein saidcompound exerts its effect on cells of both polarized and non-polarizedlineage or type, with subsequent administration or co-administration orprior administration of the modified ABC transporter polypeptide of theinvention to enhance resistance to said chemical compound in a sub-setof those cells. For example, the cytotoxic effects of a generallycytotoxic compound on the haematopoietic system of humans may bealleviated by subsequent administration, or co-administration, or prioradministration, of the modified ABC transporter polypeptide of theinvention to those haematopoietic cells, thereby enhancing theirresistance to the compound. The benefits of such an approach are evidentto those skilled in the art, particularly in so far as it relates to theapplication of cytotoxic and cytostatic compounds to cells, such as, forexample, the chemotherapeutic treatment of cancers.

[0183] The present invention extends to the use of any and all modifiedABC transporter polypeptides that are required for the influx/efflux ofa chemical compound to enhance resistance of the cell to said chemicalcompound.

[0184] As the inventive method is broadly applicable to enhancing thedrug resistance of any cell, the cell may be any polarized ornon-polarized cell or cell line referred to herein above. Preferably,the cell is a non-cancerous cell or non-infected host cell of humans orother mammals. In a particularly preferred embodiment of the invention,the cell is a non-polarized cell, such as, for example a cell of thehaematopoietic system.

[0185] The invention further extends to the use of any and all nucleicacid molecules that encode the modified ABC transporter polypeptides, toenhance the resistance of the cell to the said chemical compound.Preferably, this embodiment of the invention comprises the further stepof introducing to the cell, tissue, organ or whole organism an isolatednucleic acid that encodes the modified ABC transporter polypeptide orfunctional variant of said polypeptide.

[0186] This embodiment further includes methods of in vivo gene therapythat produce the modified ABC transporter polypeptide de novo in thecell, tissue, organ or organism, using art-recognized procedures forgene therapy. For example, bone marrow can be transduced to have analtered expression of the modified ABC transporter polypeptide, therebyconferring resistance to chemotherapeutic drugs upon bone marrow cells.Following autologous transplantation of the transduced bone marrow, amore efficient chemotherapeutic regimen can be applied to cancerpatients. The nucleic acid molecule used in performing this embodimentof the invention may be the exemplified nucleic acid described herein,or a homologue, analogue or derivative thereof encoding a modified ABCtransporter polypeptide.

[0187] The gene therapy techniques described herein can also be used toameliorate myelosuppression due to chemotherapy. In particular, theglutathione S-transferase isoenzymes having a synergistic effect withthe glutathione conjugate transporters, such as, for example, cMOAT,decrease the cytotoxicity of chemotherapeutic agents. Accordingly, oneor more vectors co-expressing the modified ABC transporter polypeptideof the invention and glutathione S-transferase are useful for increasingthe efficiency of detoxification, such as by the liver. Theco-expression of both the modified ABC transporter of the invention andglutathione S-transferase from the same or different vectors is clearlycontemplated herein.

[0188] Such gene therapy techniques can also be used to treat liverdysfunction. Liver dysfunction can result from a genetic disease (DubinJohnson's Syndrome) or due to lifestyle-influenced dysfunction resultingin cholestasis. The transplantation of non-polarized cells into liver ispossible, but these cells do not normally integrate into the structuresthat form the canalicular spaces. Moreover, the ABC transporters thatare normally distributed to the canalicular membrane of polarized cellsare localized intracellularly in such non-polarized cells. Non-polarizedcells that have been genetically transformed to express the modified ABCtransporter polypeptide of the invention function to metabolizesubstrates and transport metabolites into the sinusoidal spaces whichultimately could be filtered by the kidneys.

[0189] Additionally, the modified ABC transporter polypeptide of theinvention is used to develop novel cell lines for assaying ABCtransporter activity, substrate specificity, or drug metabolism or drugtransport. Clearly, cells expressing the modified ABC transporter of theinvention are useful in this respect for determining the role of thetransporter in the metabolism of any particular drug.

[0190] Accordingly, a further aspect of the invention contemplates asimple and reliable in vivo screening system for the discovery of novelagonists and antagonists of an ABC transporter polypeptide.

[0191] In particular, the present invention contemplates a simple andreliable in vivo screening system for discovery of novel agonists andantagonists of naturally-occurring ABC transporter polypeptides. Theobservation by the inventors that the modified ABC transporterpolypeptide of the invention has the same function as thenaturally-occurring counterpart, and is localized in the plasmamembranes of both polarized and non-polarized cell types, indicates theutility of the invention in high throughput screening to identifyagonists and antagonists of endogenous ABC transporter activities inthese cells. The present invention clearly contemplates a process whichutilizes rapid, high throughput screens with some tolerance ofnon-specificity and/or smaller-scale functional screens having higherspecificity, and/or quantitative kinetic studies to elucidate chemicalstructure/function relationships to be determined, such as, for example,the elucidation of the docking site for agonist/antagonist moleculesusing the mutants of the modified proteins.

[0192] Preferably, the present invention contemplates a process foridentifying a substrate of a native ABC transporter polypeptidecomprising:

[0193] (i) expressing the corresponding modified ABC transporterpolypeptide in a cell, wherein said modified ABC transporter polypeptideconsists of the amino acid sequence of said native ABC transporterpolypeptide wherein one or more amino acid residues of a C-terminalT-K-F motif of said native ABC transporter polypeptide having a sequenceset forth in any one of SEQ ID NOs: 52 to 58 is substituted or deleted;

[0194] (ii) determining the efflux of the compound from the cellexpressing the modified ABC transporter relative to a cell that does notexpress the native ABC transporter or the corresponding modified ABCtransporter, wherein efflux from the cell expressing the modified ABCtransporter indicates that the compound is a substrate for thecorresponding native ABC transporter.

[0195] Standard methods may be used to determine the efflux of thecompound from the cell.

[0196] In a preferred embodiment, the present invention further providesa method for identifying an inhibitor of a native ABC transporterpolypeptide comprising:

[0197] (i) expressing the corresponding modified ABC transporterpolypeptide in a cell, wherein said modified ABC transporter polypeptideconsists of the amino acid sequence of said native ABC transporterpolypeptide wherein one or more amino acid residues of a C-terminalT-K-F motif of said native ABC transporter polypeptide having a sequenceset forth in any one of SEQ ID NOs: 52 to 58 is substituted or deleted;

[0198] (ii) incubating the cell in the presence of (a) a compound beingtested for its ability to inhibit activity of the ABC transporterpolypeptide; and (b) a known substrate compound for said native ABCtransporter polypeptide;

[0199] (iii) in a separate sample to (ii), incubating the cell in thepresence of said substrate compound; and

[0200] (iv) comparing the efflux of the substrate compound at (ii) and(iii), wherein reduced efflux at (ii) compared to (iii) indicates thatthe compound being tested is an inhibitor of the native ABC transporterpolypeptide.

[0201] Based upon the similar activities of the modified ABC transporterpolypeptide and the corresponding naturally-occurring ABC transporterpolypeptide, it is preferred that the inhibitory compound identified inthis assay is also an inhibitor of the corresponding naturally-occurringABC transporter polypeptide.

[0202] It will be apparent to those skilled in the art that the assayformat described herein is readily adapted to determine novel agonistsof ABC transporter function. Accordingly, an alternative embodiment ofthis assay format provides a method for identifying an agonist of anative ABC transporter polypeptide comprising:

[0203] (i) expressing the corresponding modified ABC transporterpolypeptide in a cell, wherein said modified ABC transporter polypeptideconsists of the amino acid sequence of said native ABC transporterpolypeptide wherein one or more amino acid residues of a C-terminalT-K-F motif of said native ABC transporter polypeptide having a sequenceset forth in any one of SEQ ID NOs: 52 to 58 is substituted or deleted;

[0204] (ii) incubating the cell in the presence of (a) a compound beingtested for its ability to agonize activity of the ABC transporterpolypeptide; and (b) a known substrate compound for said native ABCtransporter polypeptide;

[0205] (iii) in a separate sample to (ii), incubating the cell in thepresence of said substrate compound; and

[0206] (iv) comparing the efflux of the substrate compound at (ii) and(iii), wherein enhanced efflux at (ii) compared to (iii) indicates thatthe compound being tested is an agonist of the native ABC transporterpolypeptide.

[0207] Based upon the similar activities of the modified ABC transporterpolypeptide and the corresponding naturally-occurring ABC transporterpolypeptide, it is preferred that the agonist identified in this assayis also an agonist of the corresponding naturally-occurring ABCtransporter polypeptide.

[0208] In an alternative embodiment, agonists may be identified by aprocess comprising:

[0209] (i) expressing a modified ABC transporter polypeptide in theplasma membrane of a polarized or non-polarized cell;

[0210] (ii) incubating the cell in the presence of (a) a compound beingtested for its ability to agonize activity of the ABC transporterpolypeptide; and (b) a known substrate compound for said modified ABCtransporter polypeptide;

[0211] (iii) in a separate sample to (ii), incubating an isogenic cellthat does not express the modified ABC transporter polypeptide in thepresence of said substrate compound and said compound being tested; and

[0212] (iv) comparing the efflux of the said substrate compound at (ii)and (iii), wherein enhanced efflux at (ii) compared to (iii) indicatesthat the compound being tested is an agonist of said modified ABCtransporter polypeptide.

[0213] Preferably, the isogenic cell does not express any ABCtransporter polypeptide capable of transporting the substrate compoundused in the assay formats described herein.

[0214] Preferred substrates which are transported by MRP1, MRP2, andMRP3 are listed in Table 4. Substrates for these transporters generallyhave a lipophilic moiety, such as, for example, bilirubin, estradiol, orarachidonate, linkes to at least one anionic residue, such as, forexample, glucuronosyl, carboxyl, glutathionyl, or sulfate. As will beknown to those skilled in the art, a conjugated substrate, particularlya glutathione conjugate, can generally be provided to the cell in anunconjugated form wherein it is conjugated by the action of anendogenous enzyme, such as, for example, glutathione-S-transferase.

[0215] Preferred substrates of modified cMOAT include leukotriene C4(LTC4; Du Pont); bilirubin; monoglucuronosyl bilirubin (Jedlitschsky etal (1997) Biochem J. 327, 305-310; Kamisako et al (1999) Hepatol. 30,485-490); bisglucuronosyl bilirubin (Jedlitschsky et al (1997) BiochemJ. 327, 305-310; Kamisako et al (1999) Hepatol. 30, 485-490);leukotriene D4 (LTD4); 1,3-chloro-2,4-dinitrobenzene; mono-chlorobimane(thiolyte, Calbiochem); 7-chloro-4-nitrobenz-2-oxa-1,3-diazole (Sigma);17β-glucuronosyl estradiol (Du Pont); 3α-sulfatolithocholyl taurine;Fluo-3 (Nies et al (1998) Hepatol. 28,1332-1340); glutathione disulphide(Leier et al (1996) Biochem J., 314, 433-437), and p-aminohippurate(Leier et al (2000) in press). For transport assays, the use of thefollowing radioligands is preferred: [³H]-LTC4 (DuPont), [³H]7β-glucuronosyl estradiol (Du Pont), [³H]monoglucuronosyl bilirubin. Theuse of the fluorescent substrate Fluo-3 is also preferred. Othersubstrates that can be readily measured include the following compoundscapable of forming glutathione conjugates:1,3-chloro-2,4-dinitrobenzene; mono-chlorobimane (thiolyte, Calbiochem);and 7-chloro-4-nitrobenz-2-oxa-1,3-diazole (Sigma). Of these compounds,1,3-chloro-2,4-dinitrobenzene is converted to DNP-SG; mono-chlorobimane(thiolyte, Calbiochem) is converted to Bimane-SG; and7-chloro-4-nitrobenz-2-oxa-1,3-diazole (Sigma) is converted to4-nitrophenyl-2-oxa-1,3-diazole-SG.

[0216] Substrates for modified MDR3 include digoxin, paclitaxel,verapamil, vinblastine, phosphatidylcholine, and short chainphosphatidylcholine analogues, and these are conveniently radiolabeledfor transport assays. for example, [12α³H]digoxin is readily availablefrom New England Nuclear Life Sciences; [³H]paclitaxel is readilyavailable from Moravek Biochemical Inc, (La Bresa, Calif., USA);[α-³²P]8-azido-ATP and [α-³²P]ATP are readily available from ICNBiomedicals (Costa Mesa, Calif., USA). [³H]verapamil has also beendescribed elsewhere as having utility in assaying for MDR3 transport(Doppenschmitt et al (1999) J Pharmacol Exp Ther 288, 348-357).

[0217] Substrates for modified MRP4 include an amphiphilic anion supra,a nucleoside analog, or cyclic nucleotide. Preferred substrates fortransport assays include the following: azidothymidine monophosphate;9-(2-phosphonylmethoxyethyl)adenine (i.e. PMEA) (Schuetz et al (1999)Nature Med 5, 1048-1051); 6-mercaptopurine;1,3-chloro-2,4-dinitrobenzene; CAMP; cGMP; Sildenafil (Pfizer);Trequinsin (Sigma); and Zaprinast (Sigma). For transport assays, thesesubstrates are conveniently provided as radiolabeled compounds. TABLE 4SUBSTRATES OF MAMMALIAN MRP POLYPEPTIDES Substrate MRP1 MRP2 MRP317β-Glucuronosyl estradiol ✓ ✓ ✓ 3α-Sulfatolithocholyltaurine ✓5-Methyltetrahydrofolate ✓ 6α-Glucuronosyl hyodeoxycholate ✓ AflatoxinB1 + reduced glutathione ✓ ampicillin ✓ Bile salt conjugates ✓Bilirubin- monoglucuronosyl- ✓ ✓ ✓ bisglucuronosyl ✓ ✓ BQ123 ✓ BQ485 ✓BQ518 ✓ bromosulfophthalein-glutathione ✓ carboxydichlorofluorescein ✓ceftriaxone ✓ Chlorambucil- ✓ monochloro-monoglutathionyl- ✓monohydroxy-monoglutathionyl- ✓ bisglutathionyl ✓ cholate3-O-glucuronide ✓ conjugated bilirubin ✓ copper (acute i.v.administration) ✓ coproporphyrin I ✓ CPT11 carboxylate ✓cysteinyl-leukotrienes ✓ Daunorubicin + reduced glutathione ✓dibromosulfophthalein ✓ dinitrophenyl-glutathione ✓ Fluo-3 ✓ ✓ Folate ✓gadolinium-ethoxybenzyl-DTPA ✓ Glucuronosyl E3040 ✓ ✓ Glucuronosyletoposide ✓ Glucuronosyl grepafloxacin ✓ Glucuronosyl nafenopin ✓Glucuronosyl SN38 carboxylate ✓ Glucuronosyl SN38 lactone ✓ glutathioneGSH ✓ glutathione GSSG ✓ Glutathione disulfide ✓ ✓glutathionyl-bromoisovalerylurea ✓ indocyanine green ✓ Leukotriene C₄ ✓✓ ✓ Leukotriene D₄ ✓ ✓ Leukotriene E₄ ✓ ✓ lithocholate 3-O-glucuronide ✓manganese ✓ Methotrexate ✓ ✓ ✓ N-Acetyl-leukotriene E₄ ✓ ✓ Melphalan-monochloro-monoglutathionyl- ✓ monohydroxy-monoglutathionyl ✓nordeoxycholate 3-O-glucuronide ✓ nordeoxycholate 3-sulfate ✓ OchratoxinA ✓ p-Aminohippurate ✓ ✓ Pravastatin ✓ S-Glutathionyl 2,4-dinitrobenzene(DNP-SG) ✓ ✓ ✓ S-Glutathionyl aflatoxin B1 ✓ S-Glutathionyl ethacrynicacid ✓ ✓ S-Glutathionyl N-ethylmaleimide ✓ S-Glutathionyl prostaglandinA₂ ✓ S-Glutathionyl sulfobromophthalein ✓ SN38 carboxylate ✓Sulfobromophthalein ✓ tauro/glycolithocholate 3-sulfate ✓taurochenodexoycholate 3-sulfate ✓ Temocaprilat ✓triiodothyronine-glucuronide ✓ Vincristine + reduced glutathione ✓ zinc✓

[0218] In accordance with the preceding embodiments, the known substratecompound used in these assays may be a cytostatic compound or cytotoxiccompound, such as, for example, any one or more of the variousantibiotics, or chemotherapeutic agents that are normally transcytosedvia an ABC transporter polypeptide from which the modified ABCtransporter polypeptide employed in the assay is derived. In adaptingthe assays to employ such cytotoxic or cytostatic compounds, enhanced orreduced efflux may be estimated by the enhanced viability and/or growthor reduced viability and/or growth, respectively, of the cell. This isbecause any enhanced efflux of the cytotoxin or cytostatic compound dueto the presence of an agonist of the modified ABC transporterpolypeptide will generally enhance cell viability and/or growth, underappropriate conditions. Similarly, any reduced efflux due to thepresence of an antagonist compound will have the effect of reducing cellsurvival and/or growth at appropriate concentrations of cytotoxin orcytostatic compound.

[0219] Preferably, the known substrate compound is capable of forming aconjugate with glutathione, glucuronate, or sulfate. For example,1-chlroro-2,4-dinitrobenzene is conjugated with glutathione to form2,4-dinitrophenylglutathione (DNP-GS). Similarly, mono-chlorobimane(thiolyte, Calbiochem) forms the glutathione conjugatebimane-glutathione. Similarly, 7-chloro-4-nitrobenz-2-oxa-1,3-diazole(Sigma) is conjugated to glutathione in the cell to form4-nitrophenyl-2-oxa-1,3-diazole glutathione. In this assay format,efflux is conveniently determined by the appearance of these substratecompounds in the media. For example, cells expressing a modified ABCtransporter are exposed briefly to 1 chloro-2,4-dinitrobenzene (CDNB),then washed and incubated with the putative agonists or antagonistsbeing tested. After incubation, the supernatant is checked byspectrophotometry for the presence of 2,4-dinitrophenyl glutathione, andits rate of appearance is a measure of the activity of the agonist orantagonist compound. This process is readily configured to a highthroughput mode (Board, P. (1984) FEBS Lett 124, 153-165; Olive andBoard (1994) Biochem. Biophys. Acta 1224, 264-268).

[0220] the assay format used may be any convenient format for assayingtransport, including a nucleotide trapping assay, or the use of cellmonolayers. Such formats are known to those skilled in the art.

[0221] In the foregoing embodiment, the use of non-polarized cells ispreferred, because they do not normally express the native counterpartof the modified ABC transporter polypeptide in their plasma membranes.However, polarized cells may also be used, because the modified ABCtransporter polypeptide accumulates over a greater surface area of theplasma membrane compared to the endogenous ABC transporter polypeptide,which is localized in the apical membrane domain. When polarized cellsare used to conduct these assays, the efflux of the cytotoxin orcytostat from the cell via the modified ABC transporter polypeptide isseveral fold (at least about 2-fold, preferably, at least about 5- to7-fold) the level of efflux via any endogenous naturally-occurring ABCtransporter polypeptide in the plasma membrane of the polarized cell.

[0222] Compounds detected using this screening procedure can ultimatelybe used, for example, as chemosensitizers in cancer therapy.

[0223] A further aspect of the invention contemplates the use of a T-K-Fmotif as a portable transport signal peptide for targeting proteins tothe apical membrane subject to the proviso that the T-K-F motif iswithin the context of an ABC transporter polypeptide.

[0224] This invention is also described with reference to the followingnon-limiting examples.

EXAMPLE 1 Modified cMOAT (MRP2) Polypeptides

[0225] Introduction

[0226] cMOAT is an ABC transporter of the subfamily known in the art asmultidrug resistance-associated proteins (MRPs). Several reviews of MRPshave been published (see Borst et al., (2000) J. Natl. Cancer Inst.92,1295-1302; and Konig et al., (1999) Biochim. Biophys. Acta 1461,377-394). There are six known human MRPs, designated MRP1 through MRP6.cMOAT corresponds to MRP2. The MRPs export a broad range of compoundsfrom the cell. MRP1 was the first and most extensively characterizedmember (Cole et al., (1992) Science 258,1650-1654) and has 49% sequenceidentity with cMOAT (Buchler et al., (1996) J. Biol. Chem. 271,15091-15098; Ito et al., (1997) Am. J. Physiol. 272, G16-G22; Paulusmaet al., (1996) Science 271,1126-1128; and Taniguchi et al., (1996)Cancer Res. 56, 4124-4129).

[0227] MRP1 and cMOAT have similar substrates, which include glutathioneconjugates, glucuronide conjugates, reduced glutathione, andchemotherapeutic drugs. The function of cMOAT was initially shown to bedistinct from MRP1 by the use of cMOAT-deficient rats GY/TR2 (Jansen etal., (1985) Hepatology 5, 573-579; Jansen et al., (1987) Hepatology 7,71-76; and Kitamura et al., (1990) Proc. Natl. Acad. Sci. U.S. A 87,3557-3561) and EHBR (Hosokawa et al., (1992) Lab. Anim. Sci. 42, 27-34).

[0228] The distribution patterns of MRP1 and cMOAT also differs. MRP1 isfound throughout the body in many tissues, including the haematopoieticsystem, the blood brain barrier, lungs, and at lower expression levelsin the liver and kidneys. In contrast, cMOAT is only found atsignificant levels in the liver and to a lesser extent in the kidneys.In these two tissues, where both proteins are expressed, they differ intheir specific cellular localization. MRP1 is found in the basolateral(sinusoidal) membrane and thus may serve to redirect potential excretionproducts back into the bloodstream. Conversely, cMOAT is solely found inthe apical membrane, and this defines its function as an export pump ofcompounds destined for terminal excretion from the body. Although bothproteins can be found in the hepatocyte, higher expression levels ofcMOAT than MRP1 create the vectorial transport of excretion productsfrom the blood into bile.

[0229] In initial experiments, haematopoietic cell lines transfectedwith cMOAT did not express a functional cMOAT due to intracellularaccumulation of the protein and minimal cell membrane localization.Similar results have since been reported by others (see Evers et al.,(1998) J. Clin. Invest 101, 1310-1319). In contrast, MRP1 shows totalcell membrane localization in similarly transfected cells.

[0230] We sought to establish the sorting signals responsible for thedifference in localization of MRP1 and cMOAT in both epithelial celllines and cells of haematopoietic lineage. Using green fluorescentprotein (GFP) fusion proteins and site-directed mutagenesis weidentified a sequence motif responsible for exclusive apicallocalization of cMOAT in polarized MDCK cells. Deletion of the motifresults in lateral localization of cMOAT in polarized MDCK cells andallows cell membrane localization in L1210 cells. The mutated proteintransports 2,4-dinitro-phenylglutathione (DNP-GS), a known substrate ofcMOAT produced by conjugation of 1-chloro-2,4-dinitrobenzene withglutathione.

[0231] Experimental Procedures

[0232] Green Fluorescent Protein (GFP) Fusion Protein Encoding GeneConstructs

[0233] GFP was fused to the C-terminal region of MRP1 or cMOATpolypeptides, to facilitate detection of the localization of theMRP1-gfp or cMOAT-gfp fusion proteins, as described below.

[0234] Human cMOAT cDNA was amplified by polymerase chain reaction usingPfuTurbo DNA polymerase (Stratagene) to remove the stop codon andintroduce restriction enzyme sites suitable for cloning. The cDNA wasamplified using a sense primer that adds an NheI site immediatelyadjacent to the start codon, as follows: (SEQ ID NO: 24)5′-AGCGCTAGCGATGCTGGAGAAGTTCTGCAAC-3′;

[0235] and an antisense primer that adds an AgeI site after the finalcodon and removes the stop codon, as follows: (SEQ ID NO: 25)5′-TACGGTACCGGTGCGAATTTTGTGCTGTTCACATTC-3′.

[0236] The polymerase chain reaction product was digested with NheI/AgeIand ligated into the NheI/AgeI-digested EGFP-N1 vector (CLONTECH).

[0237] Additionally, human MRP1 cDNA was cloned from HL60ADR cells andligated into EGFP-N1 (SaclI/AgeI) using the same polymerase chainreaction method as described in the preceding paragraphs, howeveremploying different amplification primers. The MRP1 sense primer used,which introduces a SaclI site immediately adjacent to the start codon,was as follows: (SEQ ID NO: 34) 5′-GCGGCCGCGGATGGCGCTCCGGGGCTTC-3′.

[0238] The antisense primer, which adds an AgeI site and removes thestop codon of MRP1, was as follows: (SEQ ID NO: 35)5′-TACGGTACCGGTGCCACCMGCCGGCGTCTTTGG-3′

[0239] The cMOAT-gfp and MRP1-gfp constructs supra (1 μg of DNA pertransfection) were separately transfected into MDCK cells and L1210cells using a UpofectAMINE transfection kit (Life Technologies, Inc.).Transfections of MDCK cells were carried out using Transwell plates(Costar, 24 mm×3 μm polycarbonate membrane) to enable cell polarization.Cells were imaged using a NikonTE300 inverted microscope linked to aRadiance 2000 Laser Scanning System for confocal microscopy andLasersharp 2000 imaging software (Bio-Rad).

[0240] Expression of a cMOAT Polypeptide Lacking the T-K-F motif(ΔcMOAT)

[0241] A modified cMOAT nucleotide sequence encoding a modified cMOATpolypeptide wherein the C-terminal T-K-F motif was deleted (herein“ΔcMOAT”), and without a GFP tag, was prepared using the QuikChangesite-directed mutagenesis kit. To produce this construct, template DNAcomprising the cMOAT cDNA in the mammalian expression vector pRc/CMV(Invitrogen) (Taniguchi et al., (1996) Cancer Res. 56, 4124-4129) wasamplified using a sense primer (SEQ ID NO: 26; Table 5) and antisenseprimer as follows: 5′-GGCCTTCTGCTAGCTGTTCACATTC-3′, (SEQ ID NO: 36)

[0242] thereby producing DNA wherein the nine nucleotides that encodeamino acid residues Thr1543, Lys1544, and Phe1545 of native cMOAT weredeleted.

[0243] Successful mutagenesis of two clones from separate reactions wasconfirmed by sequencing. Stable transfectants in L1210 cells wereselected for further study.

[0244] Site-Directed Mutagenesis

[0245] Substitution mutations of cMOAT were achieved using theQuikchange Site-Directed Mutagenesis Kit (Stratagene). A double-strandedplasmid vector containing the wild-type cMOAT cDNA was used as atemplate to amplify mutant sequences, using batches of syntheticcomplementary oligonucleotides (Table 5) containing the desiredmutations, which primers annealed to the 3′-end of the coding region ofthe cMOAT cDNA and were extended in a rolling circle amplificationreaction catalyzed by PfuTurbo DNA polymerase enzyme. The annealing andextension temperatures used were as recommended by the manufacturer. Inparticular, we used 18 extension cycles for 19 minutes each, to amplifyfrom 5-10 ng of template DNA in each case.

[0246] The primer sequences were thus incorporated into mutated plasmidscontaining staggered nicks. Following temperature cycling, the productwas treated with the endonuclease DpnI, to digest only the template DNAcontaining methylated and hemi-methylated sequences. The nicked vectormutant DNA was then transformed into E. coli strain XL-1 blue(Stratagene), to repair the nick and replicate the mutated DNAsequences. E. coli cells transformed with each of the mutated plasmidswas selected on kanamycin-containing plates. Colonies were cultured andDNA was isolated therefrom, and the mutations were confirmed bynucleotide sequence analysis of the recovered plasmids.

[0247] The sequences of the forward primers used in the site-directedmutagenesis of the nine nucleotides encoding the amino acid sequence ofthe C-terminal regions of several modified cMOAT polypeptides are listedin Table 5. Amino acid residues in bold type are those introduced by thesite-directed mutagenesis. The complementary nucleotide sequences of thereverse primers are readily derived. TABLE 5 Protein Sequence typeRelevant Sequence cMOAT C-terminus (SEQ ID NO: 37)    Asn Val Asn SerThr Lys Phe Primer (SEQ ID NO: 26)  G AAT GTG AAC AGC ACA AAA TTC GCCT1543A C-terminus (SEQ ID NO: 38)    Asn Val Asn Ser Ala Pro Val K1544PPrimer (SEQ ID NO: 27)  G AAT GTG AAC AGC GCA CCG GTC GCC F1545V S1542AC-terminus (SEQ ID NO: 39)    Asn Val Asn Ala Thr Lys Phe Primer (SEQ IDNO: 28)  G AAT GTG AAC GCC ACA AAA TTC GC T1543A C-terminus (SEQ ID NO:40)        Val Asn Ser Ala Lys Phe Primer (SEQ ID NO: 29)      T GTG AACAGC GCA AAA TTC GCACC K1544A C-terminus (SEQ ID NO: 41)        Val AsnSer Thr Ala Phe Primer (SEQ ID NO: 30)        GTG AAC AGC ACA GCA TTCGCACCG F1545A C-terminus (SEQ ID NO: 42)            Ser Thr Lys AlaPrimer (SEQ ID NO: 31)          C AGC ACA AAA GCC GCACCGGTCG T1543AC-terminus (SEQ ID NO: 43)    Val Asn Ser Ala Ala Ala K1544A Primer (SEQID NO: 32) AT GTG AAC AGC GCA GCA GCC GCACCGGTCC F1545A ΔT1543C-terminus (SEQ ID NO: 44)    Asn Val Asn Ser * ΔK1544 Primer (SEQ IDNO: 33)  G AAT GTG AAC AGC TAGCAGAAGGCC ΔF1545

[0248] 2,4-Dinitrophenyl Glutathione (DNP-GS) Transport

[0249] DNP-GS was generated in L1210 cells by exposure to1-chloro-2,4-dinitrobenzene and its efflux determined as describedpreviously (Olive et al., (1994) Biochim. Biophys. Acta 1224, 264-268).

[0250] Immunofluorescence

[0251] Detection and localization of untagged mutant cMOAT lacking theT-K-F motif (i.e. ΔcMOAT) was achieved by immunofluorescence, using theantibody M2 III6 (Kamiya Pty Ltd). 2×10⁵ cells were washed with PBSF(phosphate-buffered saline supplemented with 2.5% fetal bovine serum).The cells were permeabilized using digitonin (5 μg/ml) and incubated atroom temperature for 15 min. The cells were then washed three times withPBSF and then incubated with the primary antibody (2 μg) for 1 hr atroom temperature before being washed twice with PBSF. The cells wereincubated with fluorescein isothiocyanate-conjugated F(ab′)2 (Silenus,Hawthorn, Victoria, Australia) (1:80 dilution) for 30 min at roomtemperature. Finally, the cells were washed three times and resuspendedin PBSF ready for immediate confocal microscopy.

[0252] Detection of P-glycoprotein was achieved using the antibody MRK16(Kamiya Pty Ltd.). 2×10⁵ cells were washed with PBSF and incubated withthe primary antibody (2 μg) for 1 h at room temperature then washed twotimes with PBSF. The cells were incubated with fluoresceinisothiocyanate-conjugated F(ab′) (1:400 dilution) for 30 min, washedthree times, and resuspended in PBSF ready for immediate confocalmicroscopy.

[0253] Results

[0254] Localization of cMOAT gfp and MRP1-gfp Fusion Proteins in MDCKCells

[0255] To conveniently detect the localization of the proteins underinvestigation, GFP fusion proteins were produced and their localizationvisualized using confocal microscopy to visualize the fluorescentproduct, as described supra.

[0256] Using anti-cMOAT specific antibody, native cMOAT was previouslyshown to localize to the apical membrane of MDCK cells (Evers et al.,(1998) J. Clin. Invest. 101, 1310-1319; and Cul et al., (1999)Mol.Pharmacol. 55, 929-937). In the present study, human cMOAT with GFPfused to its C terminus localized to the apical membrane in polarizedMDCK cells, consistent with the localization of the native protein (FIG.1). The apical membrane of polarized MDCK cells grown on Transwellmembranes is the surface facing the media as opposed to the surfaceadhering to the membrane (basolateral).

[0257] MRP1 has been previously immune localized to the basolateralmembrane of a pig kidney epithelial cell line (LLC-PK1) (Evers et al.,(1996) J. Clin. Invest. 97,1211-1218). In the present study, human MRP1with GFP fused to its C terminus also demonstrated basolaterallocalization in polarized MDCK cells (FIG. 2).

[0258] These studies establish that fusion of MRP1 and cMOAT to GFP doesnot interfere with the normal targeting of these proteins to thebasolateral and apical membranes.

[0259] Expression of ΔcMOAT in MDCK Cells

[0260] The alignment of C-terminal sequences of MRP1 and cMOAT revealeda C-terminal motif in cMOAT (herein “T-K-F motif”) that was absent inMRP1. To determine whether the TKF motif influenced the apicallocalization of cMOAT, we constructed ΔcMOAT-gfp in which the threeC-terminal residues were deleted. When expressed in polarized MDCKcells, ΔcMOAT-gfp was found to localize predominantly in the lateraland/or basolateral membranes in contrast to the apical localization ofcMOAT-gfp (FIG. 3).

[0261] Localization of cMOAT-gfp Substitution Mutants in MDCK Cells

[0262] To further characterize the TKF motif of ΔcMOAT-gfp, and todetermine the relative importance of each residue, individual alaninemutations were introduced into each of the residue positions 1543-1545of the cMOAT-gfp construct (Table 5). Since residues 1542-1544 (S-T-K)form a predicted phosphorylation site, residue 1542 was also mutated toalanine. FIGS. 4A through 4E show the localization of each of thesemutants in MDCK cells. The effects of the substitutions were determinedby visualizing the change in localization of the mutant compared withthe native protein. The T1543A and K1544A mutants (Table 5) exhibitedboth apical and basolateral targeting with an increase in proteinaccumulation in intracellular vesicles. The F1545A mutant (Table 5) didnot exhibit modified localization in MDCK cells compared to nativecMOAT. Mutation of all three residues to alanine (i.e. the T1543A K1544AF1545A mutant in Table 5) also caused the protein to be localized to thebasolateral membrane.

[0263] Expression of cMOAT in L1210 Cells

[0264] In initial studies we found little evidence for the transport ofDNP-GS by cMOAT expressed in the mouse leukemia cell line L1210 (datanot shown). This lack of function suggests that the protein did notlocalize to the cell membrane in these cells. To confirm thisobservation, cMOAT-gfp was expressed in L1210 cells and was found tolocalize predominantly in intracellular vesicles with only minormembrane localization (FIG. 5A). Since ΔcMOAT-gfp localizedbasolaterally in MDCK cells, we were interested to determine whether itwas able to localize in the L1210 cell membrane. In contrast tocMOAT-gfp, ΔcMOAT-gfp expressed in L1210 cells almost exclusivelylocalizes to the cell membrane (FIG. 5B). To confirm the localization,we studied cells stably expressing cMOAT and ΔcMOAT without the GFP tagby immunofluorescence. cMOAT was detected intracellularly and had avesicular localization within the cell (FIG. 5C), the same distributionas shown in FIG. 5A. The ΔcMOAT polypeptide was detected in the cellmembrane (FIG. 5D), exhibiting the same localization as ΔcMOAT-gfp shownin FIG. 5B.

[0265] These data suggest that the deletion of the TKF motif from cMOATallows the successful targeting of the protein to the membrane ofnon-polarized L1210 cells. However, some L1210 cells expressing ΔcMOATor ΔcMOAT-gfp demonstrated a degree of intracellular accumulation.

[0266] 2,4-Dinitrophenyl Glutathione Transport

[0267] L1210 cells are non-adherent and non-polarized, and can bepotentially used as a convenient cell line for assessing the transportfunction of cMOAT. As shown in FIG. 6, L1210 cells stably expressingΔcMOAT showed a significantly higher efflux of DNP-GS compared tocontrol L1210 cells or L1210 cells expressing native cMOAT protein.

[0268] Discussion

[0269] Human cMOAT specifically localizes to the apical membrane ofpolarized epithelial cells in the liver and kidney. This localizationcan be replicated experimentally in MDCK cells (Evers et al., (1998) J.Clin. Invest. 101, 1310-1319; and Cui et a/, (1999) Mol. Pharmacol. 55,929-937) and LLC-PK1 cells (Chen et al., (1999) Mol. Phar-macol. 56,1219-1228; and Kawabe et al., (1999) FEBS Lett. 456, 327-331), and wedemonstrate in this study that an cMOAT-gfp fusion protein alsolocalizes to the apical membrane (FIG. 1). This allowed us to undertakemutational analysis to determine targeting signals for apicallocalization. Deletion of the three amino acids from the C terminus ofcMOAT (ΔcMOAT-gfp) caused a dramatic change in the targeting of theprotein to the basolateral membrane, and to a lesser extent in thelateral and apical membranes, in MDCK cells. The mutant's localizationin a polarized cell was similar to that of MRP1 (FIG. 2), which does notnormally have a T-K-F motif. This observation indicates that thethree-amino acid motif targets cMOAT to the apical membrane anddominates any basolateral targeting signals.

[0270] Moreover, deletion of the T-K-F motif also produces a modifiedcMOAT polypeptide that is localized in the plasma cell membrane of nonpolarized L1210 cells. In contrast, wild type cMOAT is predominantlyintracellular in L1210 cells.

[0271] To further characterize the T-K-F motif, alanine was introducedinto the position of each residue separately, and an additional mutantwas made in which all three residues were replaced by alanine.

[0272] Our data suggest that a functional T-K-F motif is characterizedby the consensus sequence S/T-X-Hy, wherein X represents any amino acidand Hy is a hydrophobic residue (Songyang et al., (1997) Science 275,73-77). For example, the T1543A mutant did exhibit modified targetingcompared with the native cMOAT protein, allowing both basolateral andapical targeting, (i.e. non-polarized targeting), and also an increasedaccumulation in vesicles, suggesting some instability in the targetingmechanism. This conclusion is also consistent with the results obtainedby the TKF-AAA mutant. The F1545A mutant did not alter normal targeting,suggesting that alanine at position 1545 is sufficiently hydrophobic fornormal targeting to occur. Accordingly, any residue (X) may be toleratedat position 1545 of cMOAT, but not at position 1544, since K1544A wasalso targeted to the basolateral membrane.

[0273] Interestingly, the serine residue at position 1542 forms apredicted phosphorylation site, and mutation of this serine residuecaused the fusion protein to localize in sub-apical vesicles. These datasuggest that S1542 may be phosphorylated and could regulate recruitmentinto the apical membrane.

[0274] There is a notable difference in the sorting of cMOAT in L1210cells compared with the MDCK cells. When expressed in L1210 cells, cMOATtransport function was minimal (results not shown). Immunofluorescencestudies of cells expressing cMOAT and confocal imaging of cellsexpressing cMOAT-gfp both confirm the intracellular localization of theprotein in L1210 cells (FIG. 5). Deletion of the T-K-F motif from cMOATand cMOAT-gfp allows the mutant protein to be expressed in the plasmamembrane, suggesting that it is this apical targeting motif thatexcludes the native cMOAT from the membrane. The stability of theprotein in the membrane also differs between MDCK cells and L1210 cells.In MDCK cells ΔcMOAT-gfp has basolateral localization in the majority ofcells.

EXAMPLE 2 TKF Motifs in Other ABC Transporter Polypeptides Derived fromSequence Alignments and Molecular Modeling

[0275] Experimental Procedure

[0276] The protein sequence of the C-terminal cytoplasmic domains of 37ABC transporters from the P-glycoprotein and MRP subfamilies werealigned with the histidine permease (HisP) sequence using the ClustalWalignment program. The multiple sequence alignment was used with thecoordinates of the HisP crystal structure (Hung et al., (1998) Nature396, 703-707) to generate a homology model of the C-terminal cytoplasmicdomain from MRP1 and cMOAT using BioNavigator at the ANGIS Internet site(BioNavigator by eBioinformatics Pty. Ltd.). The models were generatedusing the Rigorous Models software (Abagyan et al., (1994) J. Comp.Chem. 15,488-506) and presented using Swiss Pdb Viewer (v3.6b3) (Guex etal., (1997) Electrophoresis 18, 2714-2723).

[0277] Results

[0278] The alignment represented in FIG. 7 shows that those MRP proteinsthat localize to the apical membrane (cMOAT from four species) have aC-terminal T-K-F motif when compared with MRP1, MRP3, MRP5, and MRP6,which are targeted to the basolateral membrane. The P-gp, MDR3, and MRP4proteins also have a potential T-K-F motif at their C termini.

[0279] The structural coordinates for the ATP binding subunit ofhistidine permease from Salmonella typhimurium (Hung et al., (1998)Nature 396, 703-707) and the full ABC transporter sequence alignmentallowed the construction of homology models of the equivalent regions ofMRP1 and cMOAT (FIG. 8). Since the C-terminal motif of cMOAT extendsbeyond the alignment with HisP, the exact position of the T-K-F motifresidues cannot be predicted. However, the models predict that the T-K-Fmotif is positioned on the outside of the protein, away from the ATPbinding cassette and regions involved in the cytoplasmic subunitinterface. In addition, the external position of the motif would favorinteractions with other proteins involved in the targeting process.

[0280] Discussion

[0281] The deletion of the T-K-F motif increases the sequence similarityof cMOAT to MRP1 and results in the same basolateral targeting asobserved for MRP1. To investigate the tertiary structure of the subunitand the position of the motif, homology models of both MRP1 and cMOATwere created based the crystal structure of HisP. Comparisons of thehomology models clearly show the difference in length of the C terminusof MRP1 and cMOAT. It is not clear whether the TKF motif is solelyresponsible for the apical localization or whether it is the spatialarrangement of the extension and the predicted T-K-F motif that allowsbinding/modification to another part of the ABC transporter protein.From the homology model of cMOAT it appears likely that the T-K-F motifis available for interaction and not buried within the subunit Also, theposition of the C-terminus in this model suggests that the motif doesnot interact with functionally significant areas such as the ATP bindingsites. This is further supported by the ability of the deletion mutantΔcMOAT to transport 2,4-dinitrophenyl glutathione when expressed inL1210 cells (FIG. 6).

[0282] The attachment of GFP to the C-terminus of MRP1 or cMOAT did notinterfere with correct targeting of cMOAT. Based on the homology modelsin FIG. 8, the position of the C-terminal helix indicates that GFP wouldsit on the outside of the subunit. The position of GFP therefore wouldsuggest that the T-K-F motif does not need to be freely exposed andcarboxylated to function. In support of this, rabbit cMOAT has apredicted T-K-F motif in the same position as human, mouse, and ratcMOAT but also comprises a further 21 amino acids downstream (FIG. 7).

[0283] The GFP fusion proteins were expressed at consistent levels underthe CMV promoter of the EGFP-N1 vector. The cMOAT-gfp fusion proteinlocalized apically in the majority of polarized MDCK cells asrepresented in FIG. 1. cMOAT has been found to be expressed in ovariancancer cells lines (Kool et al., (1997) Cancer Res. 57, 3537-3547),renal clear cell carcinomas (Schaub et al., (1999) J. Am. Soc. Nephrol.10,1159-1169), lung, gastric, and colorectal cancer cells (Narasaki etal., (1997) Biochem. Biophys. Res. Comm. 240, 606-611).

[0284] Alignment of cMOAT with the basolateral MRP proteins MRP1, MRP3,MRP5, and MRP6 shows the absence of the motif in the basolateraltransporters (FIG. 7). Based on this alignment, the cMOAT residues1539-1545 may play a role in the targeting mechanism as this is the fulllength of the extension of the C terminus compared with the basolateralproteins.

EXAMPLE 3

[0285] Modified cMOAT Polypeptides Confer Resistance to Busulfan onL1210 Cells

[0286] Busulfan is normally conjugated to glutathione in the cytoplasmof cells by glutathione-S-transferase (Czerwinski et al. (1996) DrugMet. Dispos. 24, 1015-1019), indicating that the conjugated product ispossibly a substrate for cMOAT. Accordingly, the ability of modifiedcMOAT polypeptides to confer resistance to Busulfan was determined inL1210 cells. In particular, the ΔcMOAT polypeptide having the amino acidsequence set forth in SEQ ID NO: 4, was expressed in L1210 cells asdescribed in Example 1. The transfected cells were exposed to a range ofconcentrations of Busulfan. The survival of wild type L1210 cells, andtransfected L1210 cells expressing either native cMOAT or ΔcMOAT, wasdetermined in the presence of Busulfan. Survival was also assessedrelative to the growth of cells that had not been exposed to Busulfan.Cell growth and survival were assayed using standard procedures (Denizotet al (1986), J. Immunol. Methods 89, 271-277). Data presented in FIG. 8indicate that those cells expressing ΔcMOAT had significantly enhancedresistance to Busulfan than non-transfected L1210 cells, or L1210 cellsexpressing native cMOAT (i.e. a 2-fold increase in the IC₅₀ wasdetermined for cells expressing ΔcMOAT).

[0287] Based upon the functional equivalence of the ΔcMOAT polypeptideto the other modified cMOAT polypeptides set forth in SEQ ID NOs: 6, 10,12, and 16, those skilled in the art will be aware from this disclosureof the utility of those other sequences in conferring resistance to anychemical on a non-polarized cell.

EXAMPLE 4 Use of modified ABC Transporter Polypeptides to Screen forModulators of ABC Transporters

[0288] By targeting a modified cMOAT polypeptide to the cell membrane ofa suspension cell of the haematopoietic lineage, such as, for example,L1210 cells or Jurkat cells, therapeutic agents that are transported bycMOAT, or novel therapeutic agents that modulate cMOAT, are detected byvirtue of their ability to be transported from the cell. Cells that arestably transfected with a mutated cMOAT cDNA sequence encoding amodified cMOAT polypeptide are incubated with such novel therapeuticagents at levels that are not cytotoxic. Following incubation, thesupernatants of cells are analyzed by HPLC to determine whether or notthe agents are metabolized. Alternatively in the case of fluorescentchemical agents, the cells are examined by flow cytometry, for adecrease in fluorescence due to cMOAT export function. Using a knownfluorescent substrate for cMOAT, such as Fluo-3, potential modulators ofcMOAT are tested by detecting inhibition of the transport of thefluorescent compound, measured by flow cytometry.

[0289] Preferably, L1210 cells expressing modified ABC transporterpolypeptides (e.g. any one of the modified cMOAT polypeptides set forthin SEQ ID NOs: 4, 6, 10, 12, or 16; the modified MDR3 polypeptide of SEQID NO: 49, or the modified MRP4 polypeptide of SEQ ID NO: 51) areincubated with a suitable substrate, such as, for example,1-chloro-2,4-dinitrobenzene or mono-chlorobimane (thiolyte, Calbiochem)or 7-chloro-4-nitrobenz-2-oxa-1,3-diazole (Sigma), which are assayed bymeasuring absorbance or fluorescence. The transfected cells are thenseparately incubated with: (i) a candidate inhibitor or candidateactivator of the corresponding native ABC transporter polypeptide, beingnative cMOAT, MDR3, or MRP4, as appropriate (i.e. the test sample); and(ii) no added candidate compound (i.e. the control sample). The rate ofefflux of the glutathione conjugate from the cells is determined forboth the test sample and the control sample, by measuring the absorbanceor fluorescence of the glutathione conjugate in the medium. Thosesamples wherein the absorbance or fluorescence of the test sample issignificantly different from the absorbance or fluorescence of thecontrol sample are selected. Candidate compounds that induce higherefflux of glutathione conjugate from the cell (e.g. higher absorbance orfluorescence of the test sample relative to the absorbance orfluorescence of the control sample) are classified as agonists of thenative ABC transporter polypeptide, whilst candidate compounds thatreduce efflux of glutathione conjugate from the cell (i.e. reducedabsorbance or fluorescence of the test sample relative to the absorbanceor fluorescence of the control sample) are classified as antagonists ofthe native ABC transporter polypeptide. Optionally, this screen isreadily adapted to a high throughput format, such as, for example, byFACS screening of multiple samples, by virtue of the capability ofdetecting the glutathione conjugate.

EXAMPLE 5 A Modified MDR3 Polypeptide

[0290] Introduction

[0291] We sought to establish the localization of a modified MDR3polypeptide lacking the putative TKF motif (i.e. SEQ ID NO: 49) in bothL1210 cells and MDCK cells, using the methods established for cMOAT asdescribed in Example 1. Determination of the localization of modifiedMDR3 is facilitated by expressing the polypeptide as a green fluorescentprotein (GFP) fusion protein.

[0292] Experimental Procedures

[0293] A gene construct that encodes modified MDR3 as a fusion proteinwith GFP GFP is fused to the C-terminal region of the human MDR3polypeptide, to facilitate detection of the localization of the MDR3-gfpfusion protein, as described below.

[0294] Human MDR3 cDNA is amplified from the native MDR3-encoding cDNA(Accession No. XM 029057) by polymerase chain reaction using PfuTurboDNA polymerase (Stratagene), to remove the stop codon and introducerestriction enzyme sites suitable for cloning. DNA encoding modifiedMDR3 is amplified using a sense primer that adds an NheI siteimmediately adjacent to the start codon, as follows: (SEQ ID NO: 59)5′-AGCGCTAGCGATGGATCTTGAGGCGGCAAAG-3′;

[0295] and an antisense primer that adds an AgeI site after the finalcodon and removes the stop codon, as follows:5′-TACGGTACCGGTGCCCCAGCCTGGACA-3′; (SEQ ID NO: 60)

[0296] The polymerase chain reaction product is digested with NheI/AgeIand ligated into the NheI/AgeI-digested EGFP-N1 vector (CLONTECH), tointroduce the modified MDR3-encoding nucleotide sequence immediatelyupstream and in-frame with the GFP-encoding nucleotide sequence in thatvector.

[0297] The modified MDR3-gfp construct (1 μg of DNA per transfection) istransfected into MDCK cells and L1210 cells using a LipofectAMINEtransfection kit (Life Technologies, Inc.). Transfections of MDCK cellsare carried out using Transwell plates (Costar, 24 mm×3 μm polycarbonatemembrane) to enable cell polarization. Cells are imaged using aNikonTE300 inverted microscope linked to a Radiance 2000. Laser ScanningSystem for confocal microscopy and Lasersharp 2000 imaging software(Bio-Rad).

[0298] Expression of the Modified MDR3 Polypeptide Without a gfp Tag

[0299] The nucleotide sequence encoding the modified MDR3 polypeptide(i.e. SEQ ID NO: 48) is prepared using the QuikChange site-directedmutagenesis kit to facilitate cloning without nucleotide sequencesencoding a GFP tag. To produce this construct, template DNA comprisingthe wild-type MDR3 cDNA in the mammalian expression vector pRc/CMV(Invitrogen) (Taniguchi et al., (1996) Cancer Res. 56, 4124-4129) isamplified using primers that do not include nucleotides encoding theT-K-F motif of native MDR3. Successful mutagenesis of clones isconfirmed by sequencing, and those clones, in the pRc/CMV vector, aretransfected into L1201 cells.

[0300] The transport of [³H]paclitaxel is determined from L1210 cellsexpressing the modified MDR3 polypeptide and compared to the efflux of[³H]paclitaxel from control L1210 cells not ectopically expressing anyMDR3 polypeptide.

[0301] Results

[0302] Localization of the Modified MDR-gfp Fusion Protein

[0303] To conveniently detect the localization of the proteins underinvestigation, GFP fusion proteins are produced and their localizationis visualized using confocal microscopy to visualize the fluorescentproduct, as described supra.

[0304] When expressed in polarized MDCK cells, the modified MDR3-gfppolypeptide is found to have a modified localization compared to nativeMDR3, wherein the modified polypeptide localizes is no longerpredominantly in the apical membrane. cells.

[0305] In L1210 cells, the modified MDR3 polypeptide is found in theplasma membrane

[0306] [³H]paclitaxel Transport

[0307] L1210 cells stably expressing the modified MDR3 polypeptidewithout a GFP tag have a significantly higher efflux of [³H]paclitaxelcompared to control L1210 cells.

EXAMPLE 6 A Modified MRP4 Polypeptide

[0308] Introduction

[0309] We also sought to establish the localization of a modified MRP4polypeptide lacking the putative TKF motif (i.e. SEQ ID NO: 51), in bothL1210 cells and MDCK cells, using the methods described in Examples 1and 5, by analyzing the localization of a modified MRP4-gfp fusionpolypeptide ectopically expressed in these cell lines.

[0310] Experimental Procedures

[0311] A gene construct that encodes modified MDR3 as a fusion proteinwith GFP GFP is fused to the C-terminal region of the human MRP4polypeptide, to facilitate detection of the localization of the MRP4-gfpfusion protein, as described below.

[0312] Human MRP4 cDNA is amplified from the native MRP4-encoding cDNA(Accession No. XM 036453) by polymerase chain reaction using PfuTurboDNA polymerase (Stratagene), to remove the stop codon and introducerestriction enzyme sites suitable for cloning. The cDNA encodingmodified MRP4 is amplified using a sense primer that adds an NheI siteimmediately adjacent to the start codon, as follows: (SEQ ID NO: 61)5′-AGCGCTAGCGATGCTGCCCGTGTACCAGGAG-3′;

[0313] and an antisense primer that adds an AgeI site after the finalcodon and removes the stop codon, as follows:5′-TACGGTACCGGTGCCTCGAAAATAGTT-3′; (SEQ ID NO: 62)

[0314] The polymerase chain reaction product is digested with NheI/AgeIand ligated into the NheI/AgeI-digested EGFP-N1 vector (CLONTECH), tointroduce the modified MRP4 encoding nucleotide sequence immediatelyupstream and in-frame with the GFP-encoding nucleotide sequence in thatvector.

[0315] The modified MRP4-gfp construct (1 μl of DNA per transfection) istransfected into MDCK cells and L1210 cells using a LipofectAMINEtransfection kit (Life Technologies, Inc.). Transfections of MDCK cellsare carried out using Transwell plates (Costar, 24 mm×3 μm polycarbonatemembrane) to enable cell polarization. Cells are imaged using aNikonTE300 inverted microscope linked to a Radiance 2000 Laser ScanningSystem for confocal microscopy and Lasersharp 2000 imaging software(Bio-Rad).

[0316] Expression of the Modified MRP4 Polypeptide without a gfp Tag

[0317] The nucleotide sequence encoding the modified MRP4 polypeptide(i.e. SEQ ID NO: 50) is prepared using the QuikChange site-directedmutagenesis kit to facilitate cloning without nucleotide sequencesencoding a GFP tag. To produce this construct, template DNA comprisingthe wild-type MRP4 cDNA cloned into the mammalian expression vectorpRc/CMV (Invitrogen) (Taniguchi et al., (1996) Cancer Res. 56,4124-4129), is amplified using primers that do not include nucleotidesencoding the T-K-F motif of native MRP4. Successful mutagenesis ofclones is confirmed by sequencing, and those clones, in the pRc/CMVvector, are transfected into L1201 cells.

[0318] Radiolabeled 6-mercaptopurine is added to L1210 cells expressingthe modified MRP4 polypeptide and the efflux of 6-thio-IMP compared tothe efflux of 6-thio-IMP from L1210 cells expressing native MRP4, oralternatively, the efflux of 6-thio-IMP from control L1210 cells notectopically expressing any MRP4 polypeptide.

[0319] Results

[0320] Localization of the Modified MRP4-gfp Fusion Protein

[0321] To conveniently detect the localization of the proteins underinvestigation, GFP fusion proteins are produced and their localizationis visualized using confocal microscopy to visualize the fluorescentproduct, as described supra.

[0322] When expressed in polarized MDCK cells, the modified MRP4-gfppolypeptide is found to have a modified localization compared to nativeMRP4, wherein the modified polypeptide localizes is no longerpredominantly in the apical membrane. cells.

[0323] In L1210 cells, the modified MRP4 polypeptide is found in theplasma membrane.

[0324] 6-mercaptopurine Transport

[0325] L1210 cells stably expressing the modified MRP4 polypeptidewithout a GFP tag have a significantly higher efflux of 6-thio-IMPcompared to control L1210 cells or L1210 cells expressing native MRP4protein.

1 62 1 4635 DNA Homo sapiens CDS (1)..(4635) 1 atg ctg gag aag ttc tgcaac tct act ttt tgg aat tcc tca ttc ctg 48 Met Leu Glu Lys Phe Cys AsnSer Thr Phe Trp Asn Ser Ser Phe Leu 1 5 10 15 gac agt ccg gag gca gacctg cca ctt tgt ttt gag caa act gtt ctg 96 Asp Ser Pro Glu Ala Asp LeuPro Leu Cys Phe Glu Gln Thr Val Leu 20 25 30 gtg tgg att ccc ttg ggc ttccta tgg ctc ctg gcc ccc tgg cag ctt 144 Val Trp Ile Pro Leu Gly Phe LeuTrp Leu Leu Ala Pro Trp Gln Leu 35 40 45 ctc cac gtg tat aaa tcc agg accaag aga tcc tct acc acc aaa ctc 192 Leu His Val Tyr Lys Ser Arg Thr LysArg Ser Ser Thr Thr Lys Leu 50 55 60 tat ctt gct aag cag gta ttc gtt ggtttt ctt ctt att cta gca gcc 240 Tyr Leu Ala Lys Gln Val Phe Val Gly PheLeu Leu Ile Leu Ala Ala 65 70 75 80 ata gag ctg gcc ctt gta ctc aca gaagac tct gga caa gcc aca gtc 288 Ile Glu Leu Ala Leu Val Leu Thr Glu AspSer Gly Gln Ala Thr Val 85 90 95 cct gct gtt cga tat acc aat cca agc ctctac cta ggc aca tgg ctc 336 Pro Ala Val Arg Tyr Thr Asn Pro Ser Leu TyrLeu Gly Thr Trp Leu 100 105 110 ctg gtt ttg ctg atc caa tac agc aga caatgg tgt gta cag aaa aac 384 Leu Val Leu Leu Ile Gln Tyr Ser Arg Gln TrpCys Val Gln Lys Asn 115 120 125 tcc tgg ttc ctg tcc cta ttc tgg att ctctcg ata ctc tgt ggc act 432 Ser Trp Phe Leu Ser Leu Phe Trp Ile Leu SerIle Leu Cys Gly Thr 130 135 140 ttc caa ttt cag act ctg atc cgg aca ctctta cag ggt gac aat tct 480 Phe Gln Phe Gln Thr Leu Ile Arg Thr Leu LeuGln Gly Asp Asn Ser 145 150 155 160 aat cta gcc tac tcc tgc ctg ttc ttcatc tcc tac gga ttc cag atc 528 Asn Leu Ala Tyr Ser Cys Leu Phe Phe IleSer Tyr Gly Phe Gln Ile 165 170 175 ctg atc ctg atc ttt tca gca ttt tcagaa aat aat gag tca tca aat 576 Leu Ile Leu Ile Phe Ser Ala Phe Ser GluAsn Asn Glu Ser Ser Asn 180 185 190 aat cca tca tcc ata gct tca ttc ctgagt agc att acc tac agc tgg 624 Asn Pro Ser Ser Ile Ala Ser Phe Leu SerSer Ile Thr Tyr Ser Trp 195 200 205 tat gac agc atc att ctg aaa ggc tacaag cgt cct ctg aca ctc gag 672 Tyr Asp Ser Ile Ile Leu Lys Gly Tyr LysArg Pro Leu Thr Leu Glu 210 215 220 gat gtc tgg gaa gtt gat gaa gag atgaaa acc aag aca tta gtg agc 720 Asp Val Trp Glu Val Asp Glu Glu Met LysThr Lys Thr Leu Val Ser 225 230 235 240 aag ttt gaa acg cac atg aag agagag ctg cag aaa gcc agg cgg gca 768 Lys Phe Glu Thr His Met Lys Arg GluLeu Gln Lys Ala Arg Arg Ala 245 250 255 ctc cag aga cgg cag gag aag agctcc cag cag aac tct gga gcc agg 816 Leu Gln Arg Arg Gln Glu Lys Ser SerGln Gln Asn Ser Gly Ala Arg 260 265 270 ctg cct ggc ttg aac aag aat cagagt caa agc caa gat gcc ctt gtc 864 Leu Pro Gly Leu Asn Lys Asn Gln SerGln Ser Gln Asp Ala Leu Val 275 280 285 ctg gaa gat gtt gaa aag aaa aaaaag aag tct ggg acc aaa aaa gat 912 Leu Glu Asp Val Glu Lys Lys Lys LysLys Ser Gly Thr Lys Lys Asp 290 295 300 gtt cca aaa tcc tgg ttg atg aaggct ctg ttc aaa act ttc tac atg 960 Val Pro Lys Ser Trp Leu Met Lys AlaLeu Phe Lys Thr Phe Tyr Met 305 310 315 320 gtg ctc ctg aaa tca ttc ctactg aag cta gtg aat gac atc ttc acg 1008 Val Leu Leu Lys Ser Phe Leu LeuLys Leu Val Asn Asp Ile Phe Thr 325 330 335 ttt gtg agt cct cag ctg ctgaaa ttg ctg atc tcc ttt gca agt gac 1056 Phe Val Ser Pro Gln Leu Leu LysLeu Leu Ile Ser Phe Ala Ser Asp 340 345 350 cgt gac aca tat ttg tgg attgga tat ctc tgt gca atc ctc tta ttc 1104 Arg Asp Thr Tyr Leu Trp Ile GlyTyr Leu Cys Ala Ile Leu Leu Phe 355 360 365 act gcg gct ctc att cag tctttc tgc ctt cag tgt tat ttc caa ctg 1152 Thr Ala Ala Leu Ile Gln Ser PheCys Leu Gln Cys Tyr Phe Gln Leu 370 375 380 tgc ttc aag ctg ggt gta aaagta cgg aca gct atc atg gct tct gta 1200 Cys Phe Lys Leu Gly Val Lys ValArg Thr Ala Ile Met Ala Ser Val 385 390 395 400 tat aag aag gca ttg acccta tcc aac ttg gcc agg aag gag tac acc 1248 Tyr Lys Lys Ala Leu Thr LeuSer Asn Leu Ala Arg Lys Glu Tyr Thr 405 410 415 gtt gga gaa aca gtg aacctg atg tct gtg gat gcc cag aag ctc atg 1296 Val Gly Glu Thr Val Asn LeuMet Ser Val Asp Ala Gln Lys Leu Met 420 425 430 gat gtg acc aac ttc atgcac atg ctg tgg tca agt gtt cta cag att 1344 Asp Val Thr Asn Phe Met HisMet Leu Trp Ser Ser Val Leu Gln Ile 435 440 445 gtc tta tct atc ttc ttccta tgg aga gag ttg gga ccc tca gtc tta 1392 Val Leu Ser Ile Phe Phe LeuTrp Arg Glu Leu Gly Pro Ser Val Leu 450 455 460 gca ggt gtt ggg gtg atggtg ctt gta atc cca att aat gcg ata ctg 1440 Ala Gly Val Gly Val Met ValLeu Val Ile Pro Ile Asn Ala Ile Leu 465 470 475 480 tcc acc aag agt aagacc att cag gtc aaa aat atg aag aat aaa gac 1488 Ser Thr Lys Ser Lys ThrIle Gln Val Lys Asn Met Lys Asn Lys Asp 485 490 495 aaa cgt tta aag atcatg aat gag att ctt agt gga atc aag atc ctg 1536 Lys Arg Leu Lys Ile MetAsn Glu Ile Leu Ser Gly Ile Lys Ile Leu 500 505 510 aaa tat ttt gcc tgggaa cct tca ttc aga gac caa gta caa aac ctc 1584 Lys Tyr Phe Ala Trp GluPro Ser Phe Arg Asp Gln Val Gln Asn Leu 515 520 525 cgg aag aaa gag ctcaag aac ctg ctg gcc ttt agt caa cta cag tgt 1632 Arg Lys Lys Glu Leu LysAsn Leu Leu Ala Phe Ser Gln Leu Gln Cys 530 535 540 gta gta ata ttc gtcttc cag tta act cca gtc ctg gta tct gtg gtc 1680 Val Val Ile Phe Val PheGln Leu Thr Pro Val Leu Val Ser Val Val 545 550 555 560 aca ttt tct gtttat gtc ctg gtg gat agc aac aat att ttg gat gca 1728 Thr Phe Ser Val TyrVal Leu Val Asp Ser Asn Asn Ile Leu Asp Ala 565 570 575 caa aag gcc ttcacc tcc att acc ctc ttc aat atc ctg cgc ttt ccc 1776 Gln Lys Ala Phe ThrSer Ile Thr Leu Phe Asn Ile Leu Arg Phe Pro 580 585 590 ctg agc atg cttccc atg atg atc tcc tcc atg ctc cag gcc agt gtt 1824 Leu Ser Met Leu ProMet Met Ile Ser Ser Met Leu Gln Ala Ser Val 595 600 605 tcc aca gag cggcta gag aag tac ttg gga ggg gat gac ttg gac aca 1872 Ser Thr Glu Arg LeuGlu Lys Tyr Leu Gly Gly Asp Asp Leu Asp Thr 610 615 620 tct gcc att cgacat gac tgc aat ttt gac aaa gcc atg cag ttt tct 1920 Ser Ala Ile Arg HisAsp Cys Asn Phe Asp Lys Ala Met Gln Phe Ser 625 630 635 640 gag gcc tccttt acc tgg gaa cat gat tcg gaa gcc aca gtc cga gat 1968 Glu Ala Ser PheThr Trp Glu His Asp Ser Glu Ala Thr Val Arg Asp 645 650 655 gtg aac ctggac att atg gca ggc caa ctt gtg gct gtg ata ggc cct 2016 Val Asn Leu AspIle Met Ala Gly Gln Leu Val Ala Val Ile Gly Pro 660 665 670 gtc ggc tctggg aaa tcc tcc ttg ata tca gcc atg ctg gga gaa atg 2064 Val Gly Ser GlyLys Ser Ser Leu Ile Ser Ala Met Leu Gly Glu Met 675 680 685 gaa aat gtccac ggg cac atc acc atc aag ggc acc act gcc tat gtc 2112 Glu Asn Val HisGly His Ile Thr Ile Lys Gly Thr Thr Ala Tyr Val 690 695 700 cca cag cagtcc tgg att cag aat ggc acc ata aag gac aac atc ctt 2160 Pro Gln Gln SerTrp Ile Gln Asn Gly Thr Ile Lys Asp Asn Ile Leu 705 710 715 720 ttt ggaaca gag ttt aat gaa aag agg tac cag caa gta ctg gag gcc 2208 Phe Gly ThrGlu Phe Asn Glu Lys Arg Tyr Gln Gln Val Leu Glu Ala 725 730 735 tgt gctctc ctc cca gac ttg gaa atg ctg cct gga gga gat ttg gct 2256 Cys Ala LeuLeu Pro Asp Leu Glu Met Leu Pro Gly Gly Asp Leu Ala 740 745 750 gag attgga gag aag ggt ata aat ctt agt ggg ggt cag aag cag cgg 2304 Glu Ile GlyGlu Lys Gly Ile Asn Leu Ser Gly Gly Gln Lys Gln Arg 755 760 765 atc agcctg gcc aga gct acc tac caa aat tta gac atc tat ctt cta 2352 Ile Ser LeuAla Arg Ala Thr Tyr Gln Asn Leu Asp Ile Tyr Leu Leu 770 775 780 gat gacccc ctg tct gca gtg gat gct cat gta gga aaa cat att ttt 2400 Asp Asp ProLeu Ser Ala Val Asp Ala His Val Gly Lys His Ile Phe 785 790 795 800 aataag gtc ttg ggc ccc aat ggc ctg ttg aaa ggc aag act cga ctc 2448 Asn LysVal Leu Gly Pro Asn Gly Leu Leu Lys Gly Lys Thr Arg Leu 805 810 815 ttggtt aca cat agc atg cac ttt ctt cct caa gtg gat gag att gta 2496 Leu ValThr His Ser Met His Phe Leu Pro Gln Val Asp Glu Ile Val 820 825 830 gttctg ggg aat gga aca att gta gag aaa gga tcc tac agt gct ctc 2544 Val LeuGly Asn Gly Thr Ile Val Glu Lys Gly Ser Tyr Ser Ala Leu 835 840 845 ctggcc aaa aaa gga gag ttt gct aag aat ctg aag aca ttt cta aga 2592 Leu AlaLys Lys Gly Glu Phe Ala Lys Asn Leu Lys Thr Phe Leu Arg 850 855 860 cataca ggc cct gaa gag gaa gcc aca gtc cat gat ggc agt gaa gaa 2640 His ThrGly Pro Glu Glu Glu Ala Thr Val His Asp Gly Ser Glu Glu 865 870 875 880gaa gac gat gac tat ggg ctg ata tcc agt gtg gaa gag atc ccc gaa 2688 GluAsp Asp Asp Tyr Gly Leu Ile Ser Ser Val Glu Glu Ile Pro Glu 885 890 895gat gca gcc tcc ata acc atg aga aga gag aac agc ttt cgt cga aca 2736 AspAla Ala Ser Ile Thr Met Arg Arg Glu Asn Ser Phe Arg Arg Thr 900 905 910ctt agc cgc agt tct agg tcc aat ggc agg cat ctg aag tcc ctg aga 2784 LeuSer Arg Ser Ser Arg Ser Asn Gly Arg His Leu Lys Ser Leu Arg 915 920 925aac tcc ttg aaa act cgg aat gtg aat agc ctg aag gaa gac gaa gaa 2832 AsnSer Leu Lys Thr Arg Asn Val Asn Ser Leu Lys Glu Asp Glu Glu 930 935 940cta gtg aaa gga caa aaa cta att aag aag gaa ttc ata gaa act gga 2880 LeuVal Lys Gly Gln Lys Leu Ile Lys Lys Glu Phe Ile Glu Thr Gly 945 950 955960 aag gtg aag ttc tcc atc tac ctg gag tac cta caa gca ata gga ttg 2928Lys Val Lys Phe Ser Ile Tyr Leu Glu Tyr Leu Gln Ala Ile Gly Leu 965 970975 ttt tcg ata ttc ttc atc atc ctt gcg ttt gtg atg aat tct gtg gct 2976Phe Ser Ile Phe Phe Ile Ile Leu Ala Phe Val Met Asn Ser Val Ala 980 985990 ttt att gga tcc aac ctc tgg ctc agt gct tgg acc agt gac tct aaa 3024Phe Ile Gly Ser Asn Leu Trp Leu Ser Ala Trp Thr Ser Asp Ser Lys 995 10001005 atc ttc aat agc acc gac tat cca gca tct cag agg gac atg aga 3069Ile Phe Asn Ser Thr Asp Tyr Pro Ala Ser Gln Arg Asp Met Arg 1010 10151020 gtt gga gtc tac gga gct ctg gga tta gcc caa ggt ata ttt gtg 3114Val Gly Val Tyr Gly Ala Leu Gly Leu Ala Gln Gly Ile Phe Val 1025 10301035 ttc ata gca cat ttc tgg agt gcc ttt ggt ttc gtc cat gca tca 3159Phe Ile Ala His Phe Trp Ser Ala Phe Gly Phe Val His Ala Ser 1040 10451050 aat atc ttg cac aag caa ctg ctg aac aat atc ctt cga gca cct 3204Asn Ile Leu His Lys Gln Leu Leu Asn Asn Ile Leu Arg Ala Pro 1055 10601065 atg aga ttt ttt gac aca aca ccc aca ggc cgg att gtg aac agg 3249Met Arg Phe Phe Asp Thr Thr Pro Thr Gly Arg Ile Val Asn Arg 1070 10751080 ttt gcc ggc gat att tcc aca gtg gat gac acc ctg cct cag tcc 3294Phe Ala Gly Asp Ile Ser Thr Val Asp Asp Thr Leu Pro Gln Ser 1085 10901095 ttg cgc agc tgg att aca tgc ttc ctg ggg ata atc agc acc ctt 3339Leu Arg Ser Trp Ile Thr Cys Phe Leu Gly Ile Ile Ser Thr Leu 1100 11051110 gtc atg atc tgc atg gcc act cct gtc ttc acc atc atc gtc att 3384Val Met Ile Cys Met Ala Thr Pro Val Phe Thr Ile Ile Val Ile 1115 11201125 cct ctt ggc att att tat gta tct gtt cag atg ttt tat gtg tct 3429Pro Leu Gly Ile Ile Tyr Val Ser Val Gln Met Phe Tyr Val Ser 1130 11351140 acc tcc cgc cag ctg agg cgt ctg gac tct gtc acc agg tcc cca 3474Thr Ser Arg Gln Leu Arg Arg Leu Asp Ser Val Thr Arg Ser Pro 1145 11501155 atc tac tct cac ttc agc gag acc gta tca ggt ttg cca gtt atc 3519Ile Tyr Ser His Phe Ser Glu Thr Val Ser Gly Leu Pro Val Ile 1160 11651170 cgt gcc ttt gag cac cag cag cga ttt ctg aaa cac aat gag gag 3564Arg Ala Phe Glu His Gln Gln Arg Phe Leu Lys His Asn Glu Glu 1175 11801185 agg att gac acc aac cag aaa tgt gtc ttt tcc tgg atc acc tcc 3609Arg Ile Asp Thr Asn Gln Lys Cys Val Phe Ser Trp Ile Thr Ser 1190 11951200 aac agg tgg ctt gca att cgc ctg gag ctg gtt ggg aac ctg act 3654Asn Arg Trp Leu Ala Ile Arg Leu Glu Leu Val Gly Asn Leu Thr 1205 12101215 gtc ttc ttt tca gcc ttg atg atg gtt att tat aga gat acc cta 3699Val Phe Phe Ser Ala Leu Met Met Val Ile Tyr Arg Asp Thr Leu 1220 12251230 agt ggg gac act gtt ggc ttt gtt ctg tcc aat gca ctc aat atc 3744Ser Gly Asp Thr Val Gly Phe Val Leu Ser Asn Ala Leu Asn Ile 1235 12401245 aca caa acc ctg aac tgg ctg gtg agg atg aca tca gaa ata gag 3789Thr Gln Thr Leu Asn Trp Leu Val Arg Met Thr Ser Glu Ile Glu 1250 12551260 acc aac att gtg gct gtt gag cga ata act gag tac aca aaa gtg 3834Thr Asn Ile Val Ala Val Glu Arg Ile Thr Glu Tyr Thr Lys Val 1265 12701275 gaa aat gag gca ccc tgg gtg act gat aag agg cct ccg cca gat 3879Glu Asn Glu Ala Pro Trp Val Thr Asp Lys Arg Pro Pro Pro Asp 1280 12851290 tgg ccc agc aaa ggc aag atc cag ttt aac aac tac caa gtg cgg 3924Trp Pro Ser Lys Gly Lys Ile Gln Phe Asn Asn Tyr Gln Val Arg 1295 13001305 tac cga cct gag ctg gat ctg gtc ctc aga ggg atc act tgt gac 3969Tyr Arg Pro Glu Leu Asp Leu Val Leu Arg Gly Ile Thr Cys Asp 1310 13151320 atc ggt agc atg gag aag att ggt gtg gtg ggc agg aca gga gct 4014Ile Gly Ser Met Glu Lys Ile Gly Val Val Gly Arg Thr Gly Ala 1325 13301335 gga aag tca tcc ctc aca aac tgc ctc ttc aga atc tta gag gct 4059Gly Lys Ser Ser Leu Thr Asn Cys Leu Phe Arg Ile Leu Glu Ala 1340 13451350 gcc ggt ggt cag att atc att gat gga gta gat att gct tcc att 4104Ala Gly Gly Gln Ile Ile Ile Asp Gly Val Asp Ile Ala Ser Ile 1355 13601365 ggg ctc cac gac ctc cga gag aag ctg acc atc atc ccc cag gac 4149Gly Leu His Asp Leu Arg Glu Lys Leu Thr Ile Ile Pro Gln Asp 1370 13751380 ccc atc ctg ttc tct gga agc ctg agg atg aat ctc gac cct ttc 4194Pro Ile Leu Phe Ser Gly Ser Leu Arg Met Asn Leu Asp Pro Phe 1385 13901395 aac aac tac tca gat gag gag att tgg aag gcc ttg gag ctg gct 4239Asn Asn Tyr Ser Asp Glu Glu Ile Trp Lys Ala Leu Glu Leu Ala 1400 14051410 cac ctc aag tct ttt gtg gcc agc ctg caa ctt ggg tta tcc cac 4284His Leu Lys Ser Phe Val Ala Ser Leu Gln Leu Gly Leu Ser His 1415 14201425 gaa gtt aca gag gct ggt ggc aac ctg agc ata ggc cag agg cag 4329Glu Val Thr Glu Ala Gly Gly Asn Leu Ser Ile Gly Gln Arg Gln 1430 14351440 ctg ctg tgc ctg ggc agg gct ctg ctt cgg aaa tcc aag atc ctg 4374Leu Leu Cys Leu Gly Arg Ala Leu Leu Arg Lys Ser Lys Ile Leu 1445 14501455 gtc ctg gat gag gcc act gct gcg gtg gat cta gag aca gac aac 4419Val Leu Asp Glu Ala Thr Ala Ala Val Asp Leu Glu Thr Asp Asn 1460 14651470 ctc att cag acg acc atc caa aac gag ttc gcc cac tgc aca gtg 4464Leu Ile Gln Thr Thr Ile Gln Asn Glu Phe Ala His Cys Thr Val 1475 14801485 atc acc atc gcc cac agg ctg cat acc atc atg gac agt gac aag 4509Ile Thr Ile Ala His Arg Leu His Thr Ile Met Asp Ser Asp Lys 1490 14951500 gta atg gtc cta gac aac ggg aag att ata gag tac ggc agc cct 4554Val Met Val Leu Asp Asn Gly Lys Ile Ile Glu Tyr Gly Ser Pro 1505 15101515 gaa gaa ctg cta caa atc cct gga ccc ttt tac ttt atg gct aag 4599Glu Glu Leu Leu Gln Ile Pro Gly Pro Phe Tyr Phe Met Ala Lys 1520 15251530 gaa gct ggc att gag aat gtg aac agc aca aaa ttc 4635 Glu Ala GlyIle Glu Asn Val Asn Ser Thr Lys Phe 1535 1540 1545 2 1545 PRT Homosapiens 2 Met Leu Glu Lys Phe Cys Asn Ser Thr Phe Trp Asn Ser Ser PheLeu 1 5 10 15 Asp Ser Pro Glu Ala Asp Leu Pro Leu Cys Phe Glu Gln ThrVal Leu 20 25 30 Val Trp Ile Pro Leu Gly Phe Leu Trp Leu Leu Ala Pro TrpGln Leu 35 40 45 Leu His Val Tyr Lys Ser Arg Thr Lys Arg Ser Ser Thr ThrLys Leu 50 55 60 Tyr Leu Ala Lys Gln Val Phe Val Gly Phe Leu Leu Ile LeuAla Ala 65 70 75 80 Ile Glu Leu Ala Leu Val Leu Thr Glu Asp Ser Gly GlnAla Thr Val 85 90 95 Pro Ala Val Arg Tyr Thr Asn Pro Ser Leu Tyr Leu GlyThr Trp Leu 100 105 110 Leu Val Leu Leu Ile Gln Tyr Ser Arg Gln Trp CysVal Gln Lys Asn 115 120 125 Ser Trp Phe Leu Ser Leu Phe Trp Ile Leu SerIle Leu Cys Gly Thr 130 135 140 Phe Gln Phe Gln Thr Leu Ile Arg Thr LeuLeu Gln Gly Asp Asn Ser 145 150 155 160 Asn Leu Ala Tyr Ser Cys Leu PhePhe Ile Ser Tyr Gly Phe Gln Ile 165 170 175 Leu Ile Leu Ile Phe Ser AlaPhe Ser Glu Asn Asn Glu Ser Ser Asn 180 185 190 Asn Pro Ser Ser Ile AlaSer Phe Leu Ser Ser Ile Thr Tyr Ser Trp 195 200 205 Tyr Asp Ser Ile IleLeu Lys Gly Tyr Lys Arg Pro Leu Thr Leu Glu 210 215 220 Asp Val Trp GluVal Asp Glu Glu Met Lys Thr Lys Thr Leu Val Ser 225 230 235 240 Lys PheGlu Thr His Met Lys Arg Glu Leu Gln Lys Ala Arg Arg Ala 245 250 255 LeuGln Arg Arg Gln Glu Lys Ser Ser Gln Gln Asn Ser Gly Ala Arg 260 265 270Leu Pro Gly Leu Asn Lys Asn Gln Ser Gln Ser Gln Asp Ala Leu Val 275 280285 Leu Glu Asp Val Glu Lys Lys Lys Lys Lys Ser Gly Thr Lys Lys Asp 290295 300 Val Pro Lys Ser Trp Leu Met Lys Ala Leu Phe Lys Thr Phe Tyr Met305 310 315 320 Val Leu Leu Lys Ser Phe Leu Leu Lys Leu Val Asn Asp IlePhe Thr 325 330 335 Phe Val Ser Pro Gln Leu Leu Lys Leu Leu Ile Ser PheAla Ser Asp 340 345 350 Arg Asp Thr Tyr Leu Trp Ile Gly Tyr Leu Cys AlaIle Leu Leu Phe 355 360 365 Thr Ala Ala Leu Ile Gln Ser Phe Cys Leu GlnCys Tyr Phe Gln Leu 370 375 380 Cys Phe Lys Leu Gly Val Lys Val Arg ThrAla Ile Met Ala Ser Val 385 390 395 400 Tyr Lys Lys Ala Leu Thr Leu SerAsn Leu Ala Arg Lys Glu Tyr Thr 405 410 415 Val Gly Glu Thr Val Asn LeuMet Ser Val Asp Ala Gln Lys Leu Met 420 425 430 Asp Val Thr Asn Phe MetHis Met Leu Trp Ser Ser Val Leu Gln Ile 435 440 445 Val Leu Ser Ile PhePhe Leu Trp Arg Glu Leu Gly Pro Ser Val Leu 450 455 460 Ala Gly Val GlyVal Met Val Leu Val Ile Pro Ile Asn Ala Ile Leu 465 470 475 480 Ser ThrLys Ser Lys Thr Ile Gln Val Lys Asn Met Lys Asn Lys Asp 485 490 495 LysArg Leu Lys Ile Met Asn Glu Ile Leu Ser Gly Ile Lys Ile Leu 500 505 510Lys Tyr Phe Ala Trp Glu Pro Ser Phe Arg Asp Gln Val Gln Asn Leu 515 520525 Arg Lys Lys Glu Leu Lys Asn Leu Leu Ala Phe Ser Gln Leu Gln Cys 530535 540 Val Val Ile Phe Val Phe Gln Leu Thr Pro Val Leu Val Ser Val Val545 550 555 560 Thr Phe Ser Val Tyr Val Leu Val Asp Ser Asn Asn Ile LeuAsp Ala 565 570 575 Gln Lys Ala Phe Thr Ser Ile Thr Leu Phe Asn Ile LeuArg Phe Pro 580 585 590 Leu Ser Met Leu Pro Met Met Ile Ser Ser Met LeuGln Ala Ser Val 595 600 605 Ser Thr Glu Arg Leu Glu Lys Tyr Leu Gly GlyAsp Asp Leu Asp Thr 610 615 620 Ser Ala Ile Arg His Asp Cys Asn Phe AspLys Ala Met Gln Phe Ser 625 630 635 640 Glu Ala Ser Phe Thr Trp Glu HisAsp Ser Glu Ala Thr Val Arg Asp 645 650 655 Val Asn Leu Asp Ile Met AlaGly Gln Leu Val Ala Val Ile Gly Pro 660 665 670 Val Gly Ser Gly Lys SerSer Leu Ile Ser Ala Met Leu Gly Glu Met 675 680 685 Glu Asn Val His GlyHis Ile Thr Ile Lys Gly Thr Thr Ala Tyr Val 690 695 700 Pro Gln Gln SerTrp Ile Gln Asn Gly Thr Ile Lys Asp Asn Ile Leu 705 710 715 720 Phe GlyThr Glu Phe Asn Glu Lys Arg Tyr Gln Gln Val Leu Glu Ala 725 730 735 CysAla Leu Leu Pro Asp Leu Glu Met Leu Pro Gly Gly Asp Leu Ala 740 745 750Glu Ile Gly Glu Lys Gly Ile Asn Leu Ser Gly Gly Gln Lys Gln Arg 755 760765 Ile Ser Leu Ala Arg Ala Thr Tyr Gln Asn Leu Asp Ile Tyr Leu Leu 770775 780 Asp Asp Pro Leu Ser Ala Val Asp Ala His Val Gly Lys His Ile Phe785 790 795 800 Asn Lys Val Leu Gly Pro Asn Gly Leu Leu Lys Gly Lys ThrArg Leu 805 810 815 Leu Val Thr His Ser Met His Phe Leu Pro Gln Val AspGlu Ile Val 820 825 830 Val Leu Gly Asn Gly Thr Ile Val Glu Lys Gly SerTyr Ser Ala Leu 835 840 845 Leu Ala Lys Lys Gly Glu Phe Ala Lys Asn LeuLys Thr Phe Leu Arg 850 855 860 His Thr Gly Pro Glu Glu Glu Ala Thr ValHis Asp Gly Ser Glu Glu 865 870 875 880 Glu Asp Asp Asp Tyr Gly Leu IleSer Ser Val Glu Glu Ile Pro Glu 885 890 895 Asp Ala Ala Ser Ile Thr MetArg Arg Glu Asn Ser Phe Arg Arg Thr 900 905 910 Leu Ser Arg Ser Ser ArgSer Asn Gly Arg His Leu Lys Ser Leu Arg 915 920 925 Asn Ser Leu Lys ThrArg Asn Val Asn Ser Leu Lys Glu Asp Glu Glu 930 935 940 Leu Val Lys GlyGln Lys Leu Ile Lys Lys Glu Phe Ile Glu Thr Gly 945 950 955 960 Lys ValLys Phe Ser Ile Tyr Leu Glu Tyr Leu Gln Ala Ile Gly Leu 965 970 975 PheSer Ile Phe Phe Ile Ile Leu Ala Phe Val Met Asn Ser Val Ala 980 985 990Phe Ile Gly Ser Asn Leu Trp Leu Ser Ala Trp Thr Ser Asp Ser Lys 995 10001005 Ile Phe Asn Ser Thr Asp Tyr Pro Ala Ser Gln Arg Asp Met Arg 10101015 1020 Val Gly Val Tyr Gly Ala Leu Gly Leu Ala Gln Gly Ile Phe Val1025 1030 1035 Phe Ile Ala His Phe Trp Ser Ala Phe Gly Phe Val His AlaSer 1040 1045 1050 Asn Ile Leu His Lys Gln Leu Leu Asn Asn Ile Leu ArgAla Pro 1055 1060 1065 Met Arg Phe Phe Asp Thr Thr Pro Thr Gly Arg IleVal Asn Arg 1070 1075 1080 Phe Ala Gly Asp Ile Ser Thr Val Asp Asp ThrLeu Pro Gln Ser 1085 1090 1095 Leu Arg Ser Trp Ile Thr Cys Phe Leu GlyIle Ile Ser Thr Leu 1100 1105 1110 Val Met Ile Cys Met Ala Thr Pro ValPhe Thr Ile Ile Val Ile 1115 1120 1125 Pro Leu Gly Ile Ile Tyr Val SerVal Gln Met Phe Tyr Val Ser 1130 1135 1140 Thr Ser Arg Gln Leu Arg ArgLeu Asp Ser Val Thr Arg Ser Pro 1145 1150 1155 Ile Tyr Ser His Phe SerGlu Thr Val Ser Gly Leu Pro Val Ile 1160 1165 1170 Arg Ala Phe Glu HisGln Gln Arg Phe Leu Lys His Asn Glu Glu 1175 1180 1185 Arg Ile Asp ThrAsn Gln Lys Cys Val Phe Ser Trp Ile Thr Ser 1190 1195 1200 Asn Arg TrpLeu Ala Ile Arg Leu Glu Leu Val Gly Asn Leu Thr 1205 1210 1215 Val PhePhe Ser Ala Leu Met Met Val Ile Tyr Arg Asp Thr Leu 1220 1225 1230 SerGly Asp Thr Val Gly Phe Val Leu Ser Asn Ala Leu Asn Ile 1235 1240 1245Thr Gln Thr Leu Asn Trp Leu Val Arg Met Thr Ser Glu Ile Glu 1250 12551260 Thr Asn Ile Val Ala Val Glu Arg Ile Thr Glu Tyr Thr Lys Val 12651270 1275 Glu Asn Glu Ala Pro Trp Val Thr Asp Lys Arg Pro Pro Pro Asp1280 1285 1290 Trp Pro Ser Lys Gly Lys Ile Gln Phe Asn Asn Tyr Gln ValArg 1295 1300 1305 Tyr Arg Pro Glu Leu Asp Leu Val Leu Arg Gly Ile ThrCys Asp 1310 1315 1320 Ile Gly Ser Met Glu Lys Ile Gly Val Val Gly ArgThr Gly Ala 1325 1330 1335 Gly Lys Ser Ser Leu Thr Asn Cys Leu Phe ArgIle Leu Glu Ala 1340 1345 1350 Ala Gly Gly Gln Ile Ile Ile Asp Gly ValAsp Ile Ala Ser Ile 1355 1360 1365 Gly Leu His Asp Leu Arg Glu Lys LeuThr Ile Ile Pro Gln Asp 1370 1375 1380 Pro Ile Leu Phe Ser Gly Ser LeuArg Met Asn Leu Asp Pro Phe 1385 1390 1395 Asn Asn Tyr Ser Asp Glu GluIle Trp Lys Ala Leu Glu Leu Ala 1400 1405 1410 His Leu Lys Ser Phe ValAla Ser Leu Gln Leu Gly Leu Ser His 1415 1420 1425 Glu Val Thr Glu AlaGly Gly Asn Leu Ser Ile Gly Gln Arg Gln 1430 1435 1440 Leu Leu Cys LeuGly Arg Ala Leu Leu Arg Lys Ser Lys Ile Leu 1445 1450 1455 Val Leu AspGlu Ala Thr Ala Ala Val Asp Leu Glu Thr Asp Asn 1460 1465 1470 Leu IleGln Thr Thr Ile Gln Asn Glu Phe Ala His Cys Thr Val 1475 1480 1485 IleThr Ile Ala His Arg Leu His Thr Ile Met Asp Ser Asp Lys 1490 1495 1500Val Met Val Leu Asp Asn Gly Lys Ile Ile Glu Tyr Gly Ser Pro 1505 15101515 Glu Glu Leu Leu Gln Ile Pro Gly Pro Phe Tyr Phe Met Ala Lys 15201525 1530 Glu Ala Gly Ile Glu Asn Val Asn Ser Thr Lys Phe 1535 1540 15453 4638 DNA Homo sapiens CDS (1)..(4626) 3 atg ctg gag aag ttc tgc aactct act ttt tgg aat tcc tca ttc ctg 48 Met Leu Glu Lys Phe Cys Asn SerThr Phe Trp Asn Ser Ser Phe Leu 1 5 10 15 gac agt ccg gag gca gac ctgcca ctt tgt ttt gag caa act gtt ctg 96 Asp Ser Pro Glu Ala Asp Leu ProLeu Cys Phe Glu Gln Thr Val Leu 20 25 30 gtg tgg att ccc ttg ggc ttc ctatgg ctc ctg gcc ccc tgg cag ctt 144 Val Trp Ile Pro Leu Gly Phe Leu TrpLeu Leu Ala Pro Trp Gln Leu 35 40 45 ctc cac gtg tat aaa tcc agg acc aagaga tcc tct acc acc aaa ctc 192 Leu His Val Tyr Lys Ser Arg Thr Lys ArgSer Ser Thr Thr Lys Leu 50 55 60 tat ctt gct aag cag gta ttc gtt ggt tttctt ctt att cta gca gcc 240 Tyr Leu Ala Lys Gln Val Phe Val Gly Phe LeuLeu Ile Leu Ala Ala 65 70 75 80 ata gag ctg gcc ctt gta ctc aca gaa gactct gga caa gcc aca gtc 288 Ile Glu Leu Ala Leu Val Leu Thr Glu Asp SerGly Gln Ala Thr Val 85 90 95 cct gct gtt cga tat acc aat cca agc ctc taccta ggc aca tgg ctc 336 Pro Ala Val Arg Tyr Thr Asn Pro Ser Leu Tyr LeuGly Thr Trp Leu 100 105 110 ctg gtt ttg ctg atc caa tac agc aga caa tggtgt gta cag aaa aac 384 Leu Val Leu Leu Ile Gln Tyr Ser Arg Gln Trp CysVal Gln Lys Asn 115 120 125 tcc tgg ttc ctg tcc cta ttc tgg att ctc tcgata ctc tgt ggc act 432 Ser Trp Phe Leu Ser Leu Phe Trp Ile Leu Ser IleLeu Cys Gly Thr 130 135 140 ttc caa ttt cag act ctg atc cgg aca ctc ttacag ggt gac aat tct 480 Phe Gln Phe Gln Thr Leu Ile Arg Thr Leu Leu GlnGly Asp Asn Ser 145 150 155 160 aat cta gcc tac tcc tgc ctg ttc ttc atctcc tac gga ttc cag atc 528 Asn Leu Ala Tyr Ser Cys Leu Phe Phe Ile SerTyr Gly Phe Gln Ile 165 170 175 ctg atc ctg atc ttt tca gca ttt tca gaaaat aat gag tca tca aat 576 Leu Ile Leu Ile Phe Ser Ala Phe Ser Glu AsnAsn Glu Ser Ser Asn 180 185 190 aat cca tca tcc ata gct tca ttc ctg agtagc att acc tac agc tgg 624 Asn Pro Ser Ser Ile Ala Ser Phe Leu Ser SerIle Thr Tyr Ser Trp 195 200 205 tat gac agc atc att ctg aaa ggc tac aagcgt cct ctg aca ctc gag 672 Tyr Asp Ser Ile Ile Leu Lys Gly Tyr Lys ArgPro Leu Thr Leu Glu 210 215 220 gat gtc tgg gaa gtt gat gaa gag atg aaaacc aag aca tta gtg agc 720 Asp Val Trp Glu Val Asp Glu Glu Met Lys ThrLys Thr Leu Val Ser 225 230 235 240 aag ttt gaa acg cac atg aag aga gagctg cag aaa gcc agg cgg gca 768 Lys Phe Glu Thr His Met Lys Arg Glu LeuGln Lys Ala Arg Arg Ala 245 250 255 ctc cag aga cgg cag gag aag agc tcccag cag aac tct gga gcc agg 816 Leu Gln Arg Arg Gln Glu Lys Ser Ser GlnGln Asn Ser Gly Ala Arg 260 265 270 ctg cct ggc ttg aac aag aat cag agtcaa agc caa gat gcc ctt gtc 864 Leu Pro Gly Leu Asn Lys Asn Gln Ser GlnSer Gln Asp Ala Leu Val 275 280 285 ctg gaa gat gtt gaa aag aaa aaa aagaag tct ggg acc aaa aaa gat 912 Leu Glu Asp Val Glu Lys Lys Lys Lys LysSer Gly Thr Lys Lys Asp 290 295 300 gtt cca aaa tcc tgg ttg atg aag gctctg ttc aaa act ttc tac atg 960 Val Pro Lys Ser Trp Leu Met Lys Ala LeuPhe Lys Thr Phe Tyr Met 305 310 315 320 gtg ctc ctg aaa tca ttc cta ctgaag cta gtg aat gac atc ttc acg 1008 Val Leu Leu Lys Ser Phe Leu Leu LysLeu Val Asn Asp Ile Phe Thr 325 330 335 ttt gtg agt cct cag ctg ctg aaattg ctg atc tcc ttt gca agt gac 1056 Phe Val Ser Pro Gln Leu Leu Lys LeuLeu Ile Ser Phe Ala Ser Asp 340 345 350 cgt gac aca tat ttg tgg att ggatat ctc tgt gca atc ctc tta ttc 1104 Arg Asp Thr Tyr Leu Trp Ile Gly TyrLeu Cys Ala Ile Leu Leu Phe 355 360 365 act gcg gct ctc att cag tct ttctgc ctt cag tgt tat ttc caa ctg 1152 Thr Ala Ala Leu Ile Gln Ser Phe CysLeu Gln Cys Tyr Phe Gln Leu 370 375 380 tgc ttc aag ctg ggt gta aaa gtacgg aca gct atc atg gct tct gta 1200 Cys Phe Lys Leu Gly Val Lys Val ArgThr Ala Ile Met Ala Ser Val 385 390 395 400 tat aag aag gca ttg acc ctatcc aac ttg gcc agg aag gag tac acc 1248 Tyr Lys Lys Ala Leu Thr Leu SerAsn Leu Ala Arg Lys Glu Tyr Thr 405 410 415 gtt gga gaa aca gtg aac ctgatg tct gtg gat gcc cag aag ctc atg 1296 Val Gly Glu Thr Val Asn Leu MetSer Val Asp Ala Gln Lys Leu Met 420 425 430 gat gtg acc aac ttc atg cacatg ctg tgg tca agt gtt cta cag att 1344 Asp Val Thr Asn Phe Met His MetLeu Trp Ser Ser Val Leu Gln Ile 435 440 445 gtc tta tct atc ttc ttc ctatgg aga gag ttg gga ccc tca gtc tta 1392 Val Leu Ser Ile Phe Phe Leu TrpArg Glu Leu Gly Pro Ser Val Leu 450 455 460 gca ggt gtt ggg gtg atg gtgctt gta atc cca att aat gcg ata ctg 1440 Ala Gly Val Gly Val Met Val LeuVal Ile Pro Ile Asn Ala Ile Leu 465 470 475 480 tcc acc aag agt aag accatt cag gtc aaa aat atg aag aat aaa gac 1488 Ser Thr Lys Ser Lys Thr IleGln Val Lys Asn Met Lys Asn Lys Asp 485 490 495 aaa cgt tta aag atc atgaat gag att ctt agt gga atc aag atc ctg 1536 Lys Arg Leu Lys Ile Met AsnGlu Ile Leu Ser Gly Ile Lys Ile Leu 500 505 510 aaa tat ttt gcc tgg gaacct tca ttc aga gac caa gta caa aac ctc 1584 Lys Tyr Phe Ala Trp Glu ProSer Phe Arg Asp Gln Val Gln Asn Leu 515 520 525 cgg aag aaa gag ctc aagaac ctg ctg gcc ttt agt caa cta cag tgt 1632 Arg Lys Lys Glu Leu Lys AsnLeu Leu Ala Phe Ser Gln Leu Gln Cys 530 535 540 gta gta ata ttc gtc ttccag tta act cca gtc ctg gta tct gtg gtc 1680 Val Val Ile Phe Val Phe GlnLeu Thr Pro Val Leu Val Ser Val Val 545 550 555 560 aca ttt tct gtt tatgtc ctg gtg gat agc aac aat att ttg gat gca 1728 Thr Phe Ser Val Tyr ValLeu Val Asp Ser Asn Asn Ile Leu Asp Ala 565 570 575 caa aag gcc ttc acctcc att acc ctc ttc aat atc ctg cgc ttt ccc 1776 Gln Lys Ala Phe Thr SerIle Thr Leu Phe Asn Ile Leu Arg Phe Pro 580 585 590 ctg agc atg ctt cccatg atg atc tcc tcc atg ctc cag gcc agt gtt 1824 Leu Ser Met Leu Pro MetMet Ile Ser Ser Met Leu Gln Ala Ser Val 595 600 605 tcc aca gag cgg ctagag aag tac ttg gga ggg gat gac ttg gac aca 1872 Ser Thr Glu Arg Leu GluLys Tyr Leu Gly Gly Asp Asp Leu Asp Thr 610 615 620 tct gcc att cga catgac tgc aat ttt gac aaa gcc atg cag ttt tct 1920 Ser Ala Ile Arg His AspCys Asn Phe Asp Lys Ala Met Gln Phe Ser 625 630 635 640 gag gcc tcc tttacc tgg gaa cat gat tcg gaa gcc aca gtc cga gat 1968 Glu Ala Ser Phe ThrTrp Glu His Asp Ser Glu Ala Thr Val Arg Asp 645 650 655 gtg aac ctg gacatt atg gca ggc caa ctt gtg gct gtg ata ggc cct 2016 Val Asn Leu Asp IleMet Ala Gly Gln Leu Val Ala Val Ile Gly Pro 660 665 670 gtc ggc tct gggaaa tcc tcc ttg ata tca gcc atg ctg gga gaa atg 2064 Val Gly Ser Gly LysSer Ser Leu Ile Ser Ala Met Leu Gly Glu Met 675 680 685 gaa aat gtc cacggg cac atc acc atc aag ggc acc act gcc tat gtc 2112 Glu Asn Val His GlyHis Ile Thr Ile Lys Gly Thr Thr Ala Tyr Val 690 695 700 cca cag cag tcctgg att cag aat ggc acc ata aag gac aac atc ctt 2160 Pro Gln Gln Ser TrpIle Gln Asn Gly Thr Ile Lys Asp Asn Ile Leu 705 710 715 720 ttt gga acagag ttt aat gaa aag agg tac cag caa gta ctg gag gcc 2208 Phe Gly Thr GluPhe Asn Glu Lys Arg Tyr Gln Gln Val Leu Glu Ala 725 730 735 tgt gct ctcctc cca gac ttg gaa atg ctg cct gga gga gat ttg gct 2256 Cys Ala Leu LeuPro Asp Leu Glu Met Leu Pro Gly Gly Asp Leu Ala 740 745 750 gag att ggagag aag ggt ata aat ctt agt ggg ggt cag aag cag cgg 2304 Glu Ile Gly GluLys Gly Ile Asn Leu Ser Gly Gly Gln Lys Gln Arg 755 760 765 atc agc ctggcc aga gct acc tac caa aat tta gac atc tat ctt cta 2352 Ile Ser Leu AlaArg Ala Thr Tyr Gln Asn Leu Asp Ile Tyr Leu Leu 770 775 780 gat gac cccctg tct gca gtg gat gct cat gta gga aaa cat att ttt 2400 Asp Asp Pro LeuSer Ala Val Asp Ala His Val Gly Lys His Ile Phe 785 790 795 800 aat aaggtc ttg ggc ccc aat ggc ctg ttg aaa ggc aag act cga ctc 2448 Asn Lys ValLeu Gly Pro Asn Gly Leu Leu Lys Gly Lys Thr Arg Leu 805 810 815 ttg gttaca cat agc atg cac ttt ctt cct caa gtg gat gag att gta 2496 Leu Val ThrHis Ser Met His Phe Leu Pro Gln Val Asp Glu Ile Val 820 825 830 gtt ctgggg aat gga aca att gta gag aaa gga tcc tac agt gct ctc 2544 Val Leu GlyAsn Gly Thr Ile Val Glu Lys Gly Ser Tyr Ser Ala Leu 835 840 845 ctg gccaaa aaa gga gag ttt gct aag aat ctg aag aca ttt cta aga 2592 Leu Ala LysLys Gly Glu Phe Ala Lys Asn Leu Lys Thr Phe Leu Arg 850 855 860 cat acaggc cct gaa gag gaa gcc aca gtc cat gat ggc agt gaa gaa 2640 His Thr GlyPro Glu Glu Glu Ala Thr Val His Asp Gly Ser Glu Glu 865 870 875 880 gaagac gat gac tat ggg ctg ata tcc agt gtg gaa gag atc ccc gaa 2688 Glu AspAsp Asp Tyr Gly Leu Ile Ser Ser Val Glu Glu Ile Pro Glu 885 890 895 gatgca gcc tcc ata acc atg aga aga gag aac agc ttt cgt cga aca 2736 Asp AlaAla Ser Ile Thr Met Arg Arg Glu Asn Ser Phe Arg Arg Thr 900 905 910 cttagc cgc agt tct agg tcc aat ggc agg cat ctg aag tcc ctg aga 2784 Leu SerArg Ser Ser Arg Ser Asn Gly Arg His Leu Lys Ser Leu Arg 915 920 925 aactcc ttg aaa act cgg aat gtg aat agc ctg aag gaa gac gaa gaa 2832 Asn SerLeu Lys Thr Arg Asn Val Asn Ser Leu Lys Glu Asp Glu Glu 930 935 940 ctagtg aaa gga caa aaa cta att aag aag gaa ttc ata gaa act gga 2880 Leu ValLys Gly Gln Lys Leu Ile Lys Lys Glu Phe Ile Glu Thr Gly 945 950 955 960aag gtg aag ttc tcc atc tac ctg gag tac cta caa gca ata gga ttg 2928 LysVal Lys Phe Ser Ile Tyr Leu Glu Tyr Leu Gln Ala Ile Gly Leu 965 970 975ttt tcg ata ttc ttc atc atc ctt gcg ttt gtg atg aat tct gtg gct 2976 PheSer Ile Phe Phe Ile Ile Leu Ala Phe Val Met Asn Ser Val Ala 980 985 990ttt att gga tcc aac ctc tgg ctc agt gct tgg acc agt gac tct aaa 3024 PheIle Gly Ser Asn Leu Trp Leu Ser Ala Trp Thr Ser Asp Ser Lys 995 10001005 atc ttc aat agc acc gac tat cca gca tct cag agg gac atg aga 3069Ile Phe Asn Ser Thr Asp Tyr Pro Ala Ser Gln Arg Asp Met Arg 1010 10151020 gtt gga gtc tac gga gct ctg gga tta gcc caa ggt ata ttt gtg 3114Val Gly Val Tyr Gly Ala Leu Gly Leu Ala Gln Gly Ile Phe Val 1025 10301035 ttc ata gca cat ttc tgg agt gcc ttt ggt ttc gtc cat gca tca 3159Phe Ile Ala His Phe Trp Ser Ala Phe Gly Phe Val His Ala Ser 1040 10451050 aat atc ttg cac aag caa ctg ctg aac aat atc ctt cga gca cct 3204Asn Ile Leu His Lys Gln Leu Leu Asn Asn Ile Leu Arg Ala Pro 1055 10601065 atg aga ttt ttt gac aca aca ccc aca ggc cgg att gtg aac agg 3249Met Arg Phe Phe Asp Thr Thr Pro Thr Gly Arg Ile Val Asn Arg 1070 10751080 ttt gcc ggc gat att tcc aca gtg gat gac acc ctg cct cag tcc 3294Phe Ala Gly Asp Ile Ser Thr Val Asp Asp Thr Leu Pro Gln Ser 1085 10901095 ttg cgc agc tgg att aca tgc ttc ctg ggg ata atc agc acc ctt 3339Leu Arg Ser Trp Ile Thr Cys Phe Leu Gly Ile Ile Ser Thr Leu 1100 11051110 gtc atg atc tgc atg gcc act cct gtc ttc acc atc atc gtc att 3384Val Met Ile Cys Met Ala Thr Pro Val Phe Thr Ile Ile Val Ile 1115 11201125 cct ctt ggc att att tat gta tct gtt cag atg ttt tat gtg tct 3429Pro Leu Gly Ile Ile Tyr Val Ser Val Gln Met Phe Tyr Val Ser 1130 11351140 acc tcc cgc cag ctg agg cgt ctg gac tct gtc acc agg tcc cca 3474Thr Ser Arg Gln Leu Arg Arg Leu Asp Ser Val Thr Arg Ser Pro 1145 11501155 atc tac tct cac ttc agc gag acc gta tca ggt ttg cca gtt atc 3519Ile Tyr Ser His Phe Ser Glu Thr Val Ser Gly Leu Pro Val Ile 1160 11651170 cgt gcc ttt gag cac cag cag cga ttt ctg aaa cac aat gag gag 3564Arg Ala Phe Glu His Gln Gln Arg Phe Leu Lys His Asn Glu Glu 1175 11801185 agg att gac acc aac cag aaa tgt gtc ttt tcc tgg atc acc tcc 3609Arg Ile Asp Thr Asn Gln Lys Cys Val Phe Ser Trp Ile Thr Ser 1190 11951200 aac agg tgg ctt gca att cgc ctg gag ctg gtt ggg aac ctg act 3654Asn Arg Trp Leu Ala Ile Arg Leu Glu Leu Val Gly Asn Leu Thr 1205 12101215 gtc ttc ttt tca gcc ttg atg atg gtt att tat aga gat acc cta 3699Val Phe Phe Ser Ala Leu Met Met Val Ile Tyr Arg Asp Thr Leu 1220 12251230 agt ggg gac act gtt ggc ttt gtt ctg tcc aat gca ctc aat atc 3744Ser Gly Asp Thr Val Gly Phe Val Leu Ser Asn Ala Leu Asn Ile 1235 12401245 aca caa acc ctg aac tgg ctg gtg agg atg aca tca gaa ata gag 3789Thr Gln Thr Leu Asn Trp Leu Val Arg Met Thr Ser Glu Ile Glu 1250 12551260 acc aac att gtg gct gtt gag cga ata act gag tac aca aaa gtg 3834Thr Asn Ile Val Ala Val Glu Arg Ile Thr Glu Tyr Thr Lys Val 1265 12701275 gaa aat gag gca ccc tgg gtg act gat aag agg cct ccg cca gat 3879Glu Asn Glu Ala Pro Trp Val Thr Asp Lys Arg Pro Pro Pro Asp 1280 12851290 tgg ccc agc aaa ggc aag atc cag ttt aac aac tac caa gtg cgg 3924Trp Pro Ser Lys Gly Lys Ile Gln Phe Asn Asn Tyr Gln Val Arg 1295 13001305 tac cga cct gag ctg gat ctg gtc ctc aga ggg atc act tgt gac 3969Tyr Arg Pro Glu Leu Asp Leu Val Leu Arg Gly Ile Thr Cys Asp 1310 13151320 atc ggt agc atg gag aag att ggt gtg gtg ggc agg aca gga gct 4014Ile Gly Ser Met Glu Lys Ile Gly Val Val Gly Arg Thr Gly Ala 1325 13301335 gga aag tca tcc ctc aca aac tgc ctc ttc aga atc tta gag gct 4059Gly Lys Ser Ser Leu Thr Asn Cys Leu Phe Arg Ile Leu Glu Ala 1340 13451350 gcc ggt ggt cag att atc att gat gga gta gat att gct tcc att 4104Ala Gly Gly Gln Ile Ile Ile Asp Gly Val Asp Ile Ala Ser Ile 1355 13601365 ggg ctc cac gac ctc cga gag aag ctg acc atc atc ccc cag gac 4149Gly Leu His Asp Leu Arg Glu Lys Leu Thr Ile Ile Pro Gln Asp 1370 13751380 ccc atc ctg ttc tct gga agc ctg agg atg aat ctc gac cct ttc 4194Pro Ile Leu Phe Ser Gly Ser Leu Arg Met Asn Leu Asp Pro Phe 1385 13901395 aac aac tac tca gat gag gag att tgg aag gcc ttg gag ctg gct 4239Asn Asn Tyr Ser Asp Glu Glu Ile Trp Lys Ala Leu Glu Leu Ala 1400 14051410 cac ctc aag tct ttt gtg gcc agc ctg caa ctt ggg tta tcc cac 4284His Leu Lys Ser Phe Val Ala Ser Leu Gln Leu Gly Leu Ser His 1415 14201425 gaa gtt aca gag gct ggt ggc aac ctg agc ata ggc cag agg cag 4329Glu Val Thr Glu Ala Gly Gly Asn Leu Ser Ile Gly Gln Arg Gln 1430 14351440 ctg ctg tgc ctg ggc agg gct ctg ctt cgg aaa tcc aag atc ctg 4374Leu Leu Cys Leu Gly Arg Ala Leu Leu Arg Lys Ser Lys Ile Leu 1445 14501455 gtc ctg gat gag gcc act gct gcg gtg gat cta gag aca gac aac 4419Val Leu Asp Glu Ala Thr Ala Ala Val Asp Leu Glu Thr Asp Asn 1460 14651470 ctc att cag acg acc atc caa aac gag ttc gcc cac tgc aca gtg 4464Leu Ile Gln Thr Thr Ile Gln Asn Glu Phe Ala His Cys Thr Val 1475 14801485 atc acc atc gcc cac agg ctg cat acc atc atg gac agt gac aag 4509Ile Thr Ile Ala His Arg Leu His Thr Ile Met Asp Ser Asp Lys 1490 14951500 gta atg gtc cta gac aac ggg aag att ata gag tac ggc agc cct 4554Val Met Val Leu Asp Asn Gly Lys Ile Ile Glu Tyr Gly Ser Pro 1505 15101515 gaa gaa ctg cta caa atc cct gga ccc ttt tac ttt atg gct aag 4599Glu Glu Leu Leu Gln Ile Pro Gly Pro Phe Tyr Phe Met Ala Lys 1520 15251530 gaa gct ggc att gag aat gtg aac agc tagcagaagg cc 4638 Glu Ala GlyIle Glu Asn Val Asn Ser 1535 1540 4 1542 PRT Homo sapiens 4 Met Leu GluLys Phe Cys Asn Ser Thr Phe Trp Asn Ser Ser Phe Leu 1 5 10 15 Asp SerPro Glu Ala Asp Leu Pro Leu Cys Phe Glu Gln Thr Val Leu 20 25 30 Val TrpIle Pro Leu Gly Phe Leu Trp Leu Leu Ala Pro Trp Gln Leu 35 40 45 Leu HisVal Tyr Lys Ser Arg Thr Lys Arg Ser Ser Thr Thr Lys Leu 50 55 60 Tyr LeuAla Lys Gln Val Phe Val Gly Phe Leu Leu Ile Leu Ala Ala 65 70 75 80 IleGlu Leu Ala Leu Val Leu Thr Glu Asp Ser Gly Gln Ala Thr Val 85 90 95 ProAla Val Arg Tyr Thr Asn Pro Ser Leu Tyr Leu Gly Thr Trp Leu 100 105 110Leu Val Leu Leu Ile Gln Tyr Ser Arg Gln Trp Cys Val Gln Lys Asn 115 120125 Ser Trp Phe Leu Ser Leu Phe Trp Ile Leu Ser Ile Leu Cys Gly Thr 130135 140 Phe Gln Phe Gln Thr Leu Ile Arg Thr Leu Leu Gln Gly Asp Asn Ser145 150 155 160 Asn Leu Ala Tyr Ser Cys Leu Phe Phe Ile Ser Tyr Gly PheGln Ile 165 170 175 Leu Ile Leu Ile Phe Ser Ala Phe Ser Glu Asn Asn GluSer Ser Asn 180 185 190 Asn Pro Ser Ser Ile Ala Ser Phe Leu Ser Ser IleThr Tyr Ser Trp 195 200 205 Tyr Asp Ser Ile Ile Leu Lys Gly Tyr Lys ArgPro Leu Thr Leu Glu 210 215 220 Asp Val Trp Glu Val Asp Glu Glu Met LysThr Lys Thr Leu Val Ser 225 230 235 240 Lys Phe Glu Thr His Met Lys ArgGlu Leu Gln Lys Ala Arg Arg Ala 245 250 255 Leu Gln Arg Arg Gln Glu LysSer Ser Gln Gln Asn Ser Gly Ala Arg 260 265 270 Leu Pro Gly Leu Asn LysAsn Gln Ser Gln Ser Gln Asp Ala Leu Val 275 280 285 Leu Glu Asp Val GluLys Lys Lys Lys Lys Ser Gly Thr Lys Lys Asp 290 295 300 Val Pro Lys SerTrp Leu Met Lys Ala Leu Phe Lys Thr Phe Tyr Met 305 310 315 320 Val LeuLeu Lys Ser Phe Leu Leu Lys Leu Val Asn Asp Ile Phe Thr 325 330 335 PheVal Ser Pro Gln Leu Leu Lys Leu Leu Ile Ser Phe Ala Ser Asp 340 345 350Arg Asp Thr Tyr Leu Trp Ile Gly Tyr Leu Cys Ala Ile Leu Leu Phe 355 360365 Thr Ala Ala Leu Ile Gln Ser Phe Cys Leu Gln Cys Tyr Phe Gln Leu 370375 380 Cys Phe Lys Leu Gly Val Lys Val Arg Thr Ala Ile Met Ala Ser Val385 390 395 400 Tyr Lys Lys Ala Leu Thr Leu Ser Asn Leu Ala Arg Lys GluTyr Thr 405 410 415 Val Gly Glu Thr Val Asn Leu Met Ser Val Asp Ala GlnLys Leu Met 420 425 430 Asp Val Thr Asn Phe Met His Met Leu Trp Ser SerVal Leu Gln Ile 435 440 445 Val Leu Ser Ile Phe Phe Leu Trp Arg Glu LeuGly Pro Ser Val Leu 450 455 460 Ala Gly Val Gly Val Met Val Leu Val IlePro Ile Asn Ala Ile Leu 465 470 475 480 Ser Thr Lys Ser Lys Thr Ile GlnVal Lys Asn Met Lys Asn Lys Asp 485 490 495 Lys Arg Leu Lys Ile Met AsnGlu Ile Leu Ser Gly Ile Lys Ile Leu 500 505 510 Lys Tyr Phe Ala Trp GluPro Ser Phe Arg Asp Gln Val Gln Asn Leu 515 520 525 Arg Lys Lys Glu LeuLys Asn Leu Leu Ala Phe Ser Gln Leu Gln Cys 530 535 540 Val Val Ile PheVal Phe Gln Leu Thr Pro Val Leu Val Ser Val Val 545 550 555 560 Thr PheSer Val Tyr Val Leu Val Asp Ser Asn Asn Ile Leu Asp Ala 565 570 575 GlnLys Ala Phe Thr Ser Ile Thr Leu Phe Asn Ile Leu Arg Phe Pro 580 585 590Leu Ser Met Leu Pro Met Met Ile Ser Ser Met Leu Gln Ala Ser Val 595 600605 Ser Thr Glu Arg Leu Glu Lys Tyr Leu Gly Gly Asp Asp Leu Asp Thr 610615 620 Ser Ala Ile Arg His Asp Cys Asn Phe Asp Lys Ala Met Gln Phe Ser625 630 635 640 Glu Ala Ser Phe Thr Trp Glu His Asp Ser Glu Ala Thr ValArg Asp 645 650 655 Val Asn Leu Asp Ile Met Ala Gly Gln Leu Val Ala ValIle Gly Pro 660 665 670 Val Gly Ser Gly Lys Ser Ser Leu Ile Ser Ala MetLeu Gly Glu Met 675 680 685 Glu Asn Val His Gly His Ile Thr Ile Lys GlyThr Thr Ala Tyr Val 690 695 700 Pro Gln Gln Ser Trp Ile Gln Asn Gly ThrIle Lys Asp Asn Ile Leu 705 710 715 720 Phe Gly Thr Glu Phe Asn Glu LysArg Tyr Gln Gln Val Leu Glu Ala 725 730 735 Cys Ala Leu Leu Pro Asp LeuGlu Met Leu Pro Gly Gly Asp Leu Ala 740 745 750 Glu Ile Gly Glu Lys GlyIle Asn Leu Ser Gly Gly Gln Lys Gln Arg 755 760 765 Ile Ser Leu Ala ArgAla Thr Tyr Gln Asn Leu Asp Ile Tyr Leu Leu 770 775 780 Asp Asp Pro LeuSer Ala Val Asp Ala His Val Gly Lys His Ile Phe 785 790 795 800 Asn LysVal Leu Gly Pro Asn Gly Leu Leu Lys Gly Lys Thr Arg Leu 805 810 815 LeuVal Thr His Ser Met His Phe Leu Pro Gln Val Asp Glu Ile Val 820 825 830Val Leu Gly Asn Gly Thr Ile Val Glu Lys Gly Ser Tyr Ser Ala Leu 835 840845 Leu Ala Lys Lys Gly Glu Phe Ala Lys Asn Leu Lys Thr Phe Leu Arg 850855 860 His Thr Gly Pro Glu Glu Glu Ala Thr Val His Asp Gly Ser Glu Glu865 870 875 880 Glu Asp Asp Asp Tyr Gly Leu Ile Ser Ser Val Glu Glu IlePro Glu 885 890 895 Asp Ala Ala Ser Ile Thr Met Arg Arg Glu Asn Ser PheArg Arg Thr 900 905 910 Leu Ser Arg Ser Ser Arg Ser Asn Gly Arg His LeuLys Ser Leu Arg 915 920 925 Asn Ser Leu Lys Thr Arg Asn Val Asn Ser LeuLys Glu Asp Glu Glu 930 935 940 Leu Val Lys Gly Gln Lys Leu Ile Lys LysGlu Phe Ile Glu Thr Gly 945 950 955 960 Lys Val Lys Phe Ser Ile Tyr LeuGlu Tyr Leu Gln Ala Ile Gly Leu 965 970 975 Phe Ser Ile Phe Phe Ile IleLeu Ala Phe Val Met Asn Ser Val Ala 980 985 990 Phe Ile Gly Ser Asn LeuTrp Leu Ser Ala Trp Thr Ser Asp Ser Lys 995 1000 1005 Ile Phe Asn SerThr Asp Tyr Pro Ala Ser Gln Arg Asp Met Arg 1010 1015 1020 Val Gly ValTyr Gly Ala Leu Gly Leu Ala Gln Gly Ile Phe Val 1025 1030 1035 Phe IleAla His Phe Trp Ser Ala Phe Gly Phe Val His Ala Ser 1040 1045 1050 AsnIle Leu His Lys Gln Leu Leu Asn Asn Ile Leu Arg Ala Pro 1055 1060 1065Met Arg Phe Phe Asp Thr Thr Pro Thr Gly Arg Ile Val Asn Arg 1070 10751080 Phe Ala Gly Asp Ile Ser Thr Val Asp Asp Thr Leu Pro Gln Ser 10851090 1095 Leu Arg Ser Trp Ile Thr Cys Phe Leu Gly Ile Ile Ser Thr Leu1100 1105 1110 Val Met Ile Cys Met Ala Thr Pro Val Phe Thr Ile Ile ValIle 1115 1120 1125 Pro Leu Gly Ile Ile Tyr Val Ser Val Gln Met Phe TyrVal Ser 1130 1135 1140 Thr Ser Arg Gln Leu Arg Arg Leu Asp Ser Val ThrArg Ser Pro 1145 1150 1155 Ile Tyr Ser His Phe Ser Glu Thr Val Ser GlyLeu Pro Val Ile 1160 1165 1170 Arg Ala Phe Glu His Gln Gln Arg Phe LeuLys His Asn Glu Glu 1175 1180 1185 Arg Ile Asp Thr Asn Gln Lys Cys ValPhe Ser Trp Ile Thr Ser 1190 1195 1200 Asn Arg Trp Leu Ala Ile Arg LeuGlu Leu Val Gly Asn Leu Thr 1205 1210 1215 Val Phe Phe Ser Ala Leu MetMet Val Ile Tyr Arg Asp Thr Leu 1220 1225 1230 Ser Gly Asp Thr Val GlyPhe Val Leu Ser Asn Ala Leu Asn Ile 1235 1240 1245 Thr Gln Thr Leu AsnTrp Leu Val Arg Met Thr Ser Glu Ile Glu 1250 1255 1260 Thr Asn Ile ValAla Val Glu Arg Ile Thr Glu Tyr Thr Lys Val 1265 1270 1275 Glu Asn GluAla Pro Trp Val Thr Asp Lys Arg Pro Pro Pro Asp 1280 1285 1290 Trp ProSer Lys Gly Lys Ile Gln Phe Asn Asn Tyr Gln Val Arg 1295 1300 1305 TyrArg Pro Glu Leu Asp Leu Val Leu Arg Gly Ile Thr Cys Asp 1310 1315 1320Ile Gly Ser Met Glu Lys Ile Gly Val Val Gly Arg Thr Gly Ala 1325 13301335 Gly Lys Ser Ser Leu Thr Asn Cys Leu Phe Arg Ile Leu Glu Ala 13401345 1350 Ala Gly Gly Gln Ile Ile Ile Asp Gly Val Asp Ile Ala Ser Ile1355 1360 1365 Gly Leu His Asp Leu Arg Glu Lys Leu Thr Ile Ile Pro GlnAsp 1370 1375 1380 Pro Ile Leu Phe Ser Gly Ser Leu Arg Met Asn Leu AspPro Phe 1385 1390 1395 Asn Asn Tyr Ser Asp Glu Glu Ile Trp Lys Ala LeuGlu Leu Ala 1400 1405 1410 His Leu Lys Ser Phe Val Ala Ser Leu Gln LeuGly Leu Ser His 1415 1420 1425 Glu Val Thr Glu Ala Gly Gly Asn Leu SerIle Gly Gln Arg Gln 1430 1435 1440 Leu Leu Cys Leu Gly Arg Ala Leu LeuArg Lys Ser Lys Ile Leu 1445 1450 1455 Val Leu Asp Glu Ala Thr Ala AlaVal Asp Leu Glu Thr Asp Asn 1460 1465 1470 Leu Ile Gln Thr Thr Ile GlnAsn Glu Phe Ala His Cys Thr Val 1475 1480 1485 Ile Thr Ile Ala His ArgLeu His Thr Ile Met Asp Ser Asp Lys 1490 1495 1500 Val Met Val Leu AspAsn Gly Lys Ile Ile Glu Tyr Gly Ser Pro 1505 1510 1515 Glu Glu Leu LeuGln Ile Pro Gly Pro Phe Tyr Phe Met Ala Lys 1520 1525 1530 Glu Ala GlyIle Glu Asn Val Asn Ser 1535 1540 5 4638 DNA Homo sapiens CDS(1)..(4635) 5 atg ctg gag aag ttc tgc aac tct act ttt tgg aat tcc tcattc ctg 48 Met Leu Glu Lys Phe Cys Asn Ser Thr Phe Trp Asn Ser Ser PheLeu 1 5 10 15 gac agt ccg gag gca gac ctg cca ctt tgt ttt gag caa actgtt ctg 96 Asp Ser Pro Glu Ala Asp Leu Pro Leu Cys Phe Glu Gln Thr ValLeu 20 25 30 gtg tgg att ccc ttg ggc ttc cta tgg ctc ctg gcc ccc tgg cagctt 144 Val Trp Ile Pro Leu Gly Phe Leu Trp Leu Leu Ala Pro Trp Gln Leu35 40 45 ctc cac gtg tat aaa tcc agg acc aag aga tcc tct acc acc aaa ctc192 Leu His Val Tyr Lys Ser Arg Thr Lys Arg Ser Ser Thr Thr Lys Leu 5055 60 tat ctt gct aag cag gta ttc gtt ggt ttt ctt ctt att cta gca gcc240 Tyr Leu Ala Lys Gln Val Phe Val Gly Phe Leu Leu Ile Leu Ala Ala 6570 75 80 ata gag ctg gcc ctt gta ctc aca gaa gac tct gga caa gcc aca gtc288 Ile Glu Leu Ala Leu Val Leu Thr Glu Asp Ser Gly Gln Ala Thr Val 8590 95 cct gct gtt cga tat acc aat cca agc ctc tac cta ggc aca tgg ctc336 Pro Ala Val Arg Tyr Thr Asn Pro Ser Leu Tyr Leu Gly Thr Trp Leu 100105 110 ctg gtt ttg ctg atc caa tac agc aga caa tgg tgt gta cag aaa aac384 Leu Val Leu Leu Ile Gln Tyr Ser Arg Gln Trp Cys Val Gln Lys Asn 115120 125 tcc tgg ttc ctg tcc cta ttc tgg att ctc tcg ata ctc tgt ggc act432 Ser Trp Phe Leu Ser Leu Phe Trp Ile Leu Ser Ile Leu Cys Gly Thr 130135 140 ttc caa ttt cag act ctg atc cgg aca ctc tta cag ggt gac aat tct480 Phe Gln Phe Gln Thr Leu Ile Arg Thr Leu Leu Gln Gly Asp Asn Ser 145150 155 160 aat cta gcc tac tcc tgc ctg ttc ttc atc tcc tac gga ttc cagatc 528 Asn Leu Ala Tyr Ser Cys Leu Phe Phe Ile Ser Tyr Gly Phe Gln Ile165 170 175 ctg atc ctg atc ttt tca gca ttt tca gaa aat aat gag tca tcaaat 576 Leu Ile Leu Ile Phe Ser Ala Phe Ser Glu Asn Asn Glu Ser Ser Asn180 185 190 aat cca tca tcc ata gct tca ttc ctg agt agc att acc tac agctgg 624 Asn Pro Ser Ser Ile Ala Ser Phe Leu Ser Ser Ile Thr Tyr Ser Trp195 200 205 tat gac agc atc att ctg aaa ggc tac aag cgt cct ctg aca ctcgag 672 Tyr Asp Ser Ile Ile Leu Lys Gly Tyr Lys Arg Pro Leu Thr Leu Glu210 215 220 gat gtc tgg gaa gtt gat gaa gag atg aaa acc aag aca tta gtgagc 720 Asp Val Trp Glu Val Asp Glu Glu Met Lys Thr Lys Thr Leu Val Ser225 230 235 240 aag ttt gaa acg cac atg aag aga gag ctg cag aaa gcc aggcgg gca 768 Lys Phe Glu Thr His Met Lys Arg Glu Leu Gln Lys Ala Arg ArgAla 245 250 255 ctc cag aga cgg cag gag aag agc tcc cag cag aac tct ggagcc agg 816 Leu Gln Arg Arg Gln Glu Lys Ser Ser Gln Gln Asn Ser Gly AlaArg 260 265 270 ctg cct ggc ttg aac aag aat cag agt caa agc caa gat gccctt gtc 864 Leu Pro Gly Leu Asn Lys Asn Gln Ser Gln Ser Gln Asp Ala LeuVal 275 280 285 ctg gaa gat gtt gaa aag aaa aaa aag aag tct ggg acc aaaaaa gat 912 Leu Glu Asp Val Glu Lys Lys Lys Lys Lys Ser Gly Thr Lys LysAsp 290 295 300 gtt cca aaa tcc tgg ttg atg aag gct ctg ttc aaa act ttctac atg 960 Val Pro Lys Ser Trp Leu Met Lys Ala Leu Phe Lys Thr Phe TyrMet 305 310 315 320 gtg ctc ctg aaa tca ttc cta ctg aag cta gtg aat gacatc ttc acg 1008 Val Leu Leu Lys Ser Phe Leu Leu Lys Leu Val Asn Asp IlePhe Thr 325 330 335 ttt gtg agt cct cag ctg ctg aaa ttg ctg atc tcc tttgca agt gac 1056 Phe Val Ser Pro Gln Leu Leu Lys Leu Leu Ile Ser Phe AlaSer Asp 340 345 350 cgt gac aca tat ttg tgg att gga tat ctc tgt gca atcctc tta ttc 1104 Arg Asp Thr Tyr Leu Trp Ile Gly Tyr Leu Cys Ala Ile LeuLeu Phe 355 360 365 act gcg gct ctc att cag tct ttc tgc ctt cag tgt tatttc caa ctg 1152 Thr Ala Ala Leu Ile Gln Ser Phe Cys Leu Gln Cys Tyr PheGln Leu 370 375 380 tgc ttc aag ctg ggt gta aaa gta cgg aca gct atc atggct tct gta 1200 Cys Phe Lys Leu Gly Val Lys Val Arg Thr Ala Ile Met AlaSer Val 385 390 395 400 tat aag aag gca ttg acc cta tcc aac ttg gcc aggaag gag tac acc 1248 Tyr Lys Lys Ala Leu Thr Leu Ser Asn Leu Ala Arg LysGlu Tyr Thr 405 410 415 gtt gga gaa aca gtg aac ctg atg tct gtg gat gcccag aag ctc atg 1296 Val Gly Glu Thr Val Asn Leu Met Ser Val Asp Ala GlnLys Leu Met 420 425 430 gat gtg acc aac ttc atg cac atg ctg tgg tca agtgtt cta cag att 1344 Asp Val Thr Asn Phe Met His Met Leu Trp Ser Ser ValLeu Gln Ile 435 440 445 gtc tta tct atc ttc ttc cta tgg aga gag ttg ggaccc tca gtc tta 1392 Val Leu Ser Ile Phe Phe Leu Trp Arg Glu Leu Gly ProSer Val Leu 450 455 460 gca ggt gtt ggg gtg atg gtg ctt gta atc cca attaat gcg ata ctg 1440 Ala Gly Val Gly Val Met Val Leu Val Ile Pro Ile AsnAla Ile Leu 465 470 475 480 tcc acc aag agt aag acc att cag gtc aaa aatatg aag aat aaa gac 1488 Ser Thr Lys Ser Lys Thr Ile Gln Val Lys Asn MetLys Asn Lys Asp 485 490 495 aaa cgt tta aag atc atg aat gag att ctt agtgga atc aag atc ctg 1536 Lys Arg Leu Lys Ile Met Asn Glu Ile Leu Ser GlyIle Lys Ile Leu 500 505 510 aaa tat ttt gcc tgg gaa cct tca ttc aga gaccaa gta caa aac ctc 1584 Lys Tyr Phe Ala Trp Glu Pro Ser Phe Arg Asp GlnVal Gln Asn Leu 515 520 525 cgg aag aaa gag ctc aag aac ctg ctg gcc tttagt caa cta cag tgt 1632 Arg Lys Lys Glu Leu Lys Asn Leu Leu Ala Phe SerGln Leu Gln Cys 530 535 540 gta gta ata ttc gtc ttc cag tta act cca gtcctg gta tct gtg gtc 1680 Val Val Ile Phe Val Phe Gln Leu Thr Pro Val LeuVal Ser Val Val 545 550 555 560 aca ttt tct gtt tat gtc ctg gtg gat agcaac aat att ttg gat gca 1728 Thr Phe Ser Val Tyr Val Leu Val Asp Ser AsnAsn Ile Leu Asp Ala 565 570 575 caa aag gcc ttc acc tcc att acc ctc ttcaat atc ctg cgc ttt ccc 1776 Gln Lys Ala Phe Thr Ser Ile Thr Leu Phe AsnIle Leu Arg Phe Pro 580 585 590 ctg agc atg ctt ccc atg atg atc tcc tccatg ctc cag gcc agt gtt 1824 Leu Ser Met Leu Pro Met Met Ile Ser Ser MetLeu Gln Ala Ser Val 595 600 605 tcc aca gag cgg cta gag aag tac ttg ggaggg gat gac ttg gac aca 1872 Ser Thr Glu Arg Leu Glu Lys Tyr Leu Gly GlyAsp Asp Leu Asp Thr 610 615 620 tct gcc att cga cat gac tgc aat ttt gacaaa gcc atg cag ttt tct 1920 Ser Ala Ile Arg His Asp Cys Asn Phe Asp LysAla Met Gln Phe Ser 625 630 635 640 gag gcc tcc ttt acc tgg gaa cat gattcg gaa gcc aca gtc cga gat 1968 Glu Ala Ser Phe Thr Trp Glu His Asp SerGlu Ala Thr Val Arg Asp 645 650 655 gtg aac ctg gac att atg gca ggc caactt gtg gct gtg ata ggc cct 2016 Val Asn Leu Asp Ile Met Ala Gly Gln LeuVal Ala Val Ile Gly Pro 660 665 670 gtc ggc tct ggg aaa tcc tcc ttg atatca gcc atg ctg gga gaa atg 2064 Val Gly Ser Gly Lys Ser Ser Leu Ile SerAla Met Leu Gly Glu Met 675 680 685 gaa aat gtc cac ggg cac atc acc atcaag ggc acc act gcc tat gtc 2112 Glu Asn Val His Gly His Ile Thr Ile LysGly Thr Thr Ala Tyr Val 690 695 700 cca cag cag tcc tgg att cag aat ggcacc ata aag gac aac atc ctt 2160 Pro Gln Gln Ser Trp Ile Gln Asn Gly ThrIle Lys Asp Asn Ile Leu 705 710 715 720 ttt gga aca gag ttt aat gaa aagagg tac cag caa gta ctg gag gcc 2208 Phe Gly Thr Glu Phe Asn Glu Lys ArgTyr Gln Gln Val Leu Glu Ala 725 730 735 tgt gct ctc ctc cca gac ttg gaaatg ctg cct gga gga gat ttg gct 2256 Cys Ala Leu Leu Pro Asp Leu Glu MetLeu Pro Gly Gly Asp Leu Ala 740 745 750 gag att gga gag aag ggt ata aatctt agt ggg ggt cag aag cag cgg 2304 Glu Ile Gly Glu Lys Gly Ile Asn LeuSer Gly Gly Gln Lys Gln Arg 755 760 765 atc agc ctg gcc aga gct acc taccaa aat tta gac atc tat ctt cta 2352 Ile Ser Leu Ala Arg Ala Thr Tyr GlnAsn Leu Asp Ile Tyr Leu Leu 770 775 780 gat gac ccc ctg tct gca gtg gatgct cat gta gga aaa cat att ttt 2400 Asp Asp Pro Leu Ser Ala Val Asp AlaHis Val Gly Lys His Ile Phe 785 790 795 800 aat aag gtc ttg ggc ccc aatggc ctg ttg aaa ggc aag act cga ctc 2448 Asn Lys Val Leu Gly Pro Asn GlyLeu Leu Lys Gly Lys Thr Arg Leu 805 810 815 ttg gtt aca cat agc atg cacttt ctt cct caa gtg gat gag att gta 2496 Leu Val Thr His Ser Met His PheLeu Pro Gln Val Asp Glu Ile Val 820 825 830 gtt ctg ggg aat gga aca attgta gag aaa gga tcc tac agt gct ctc 2544 Val Leu Gly Asn Gly Thr Ile ValGlu Lys Gly Ser Tyr Ser Ala Leu 835 840 845 ctg gcc aaa aaa gga gag tttgct aag aat ctg aag aca ttt cta aga 2592 Leu Ala Lys Lys Gly Glu Phe AlaLys Asn Leu Lys Thr Phe Leu Arg 850 855 860 cat aca ggc cct gaa gag gaagcc aca gtc cat gat ggc agt gaa gaa 2640 His Thr Gly Pro Glu Glu Glu AlaThr Val His Asp Gly Ser Glu Glu 865 870 875 880 gaa gac gat gac tat gggctg ata tcc agt gtg gaa gag atc ccc gaa 2688 Glu Asp Asp Asp Tyr Gly LeuIle Ser Ser Val Glu Glu Ile Pro Glu 885 890 895 gat gca gcc tcc ata accatg aga aga gag aac agc ttt cgt cga aca 2736 Asp Ala Ala Ser Ile Thr MetArg Arg Glu Asn Ser Phe Arg Arg Thr 900 905 910 ctt agc cgc agt tct aggtcc aat ggc agg cat ctg aag tcc ctg aga 2784 Leu Ser Arg Ser Ser Arg SerAsn Gly Arg His Leu Lys Ser Leu Arg 915 920 925 aac tcc ttg aaa act cggaat gtg aat agc ctg aag gaa gac gaa gaa 2832 Asn Ser Leu Lys Thr Arg AsnVal Asn Ser Leu Lys Glu Asp Glu Glu 930 935 940 cta gtg aaa gga caa aaacta att aag aag gaa ttc ata gaa act gga 2880 Leu Val Lys Gly Gln Lys LeuIle Lys Lys Glu Phe Ile Glu Thr Gly 945 950 955 960 aag gtg aag ttc tccatc tac ctg gag tac cta caa gca ata gga ttg 2928 Lys Val Lys Phe Ser IleTyr Leu Glu Tyr Leu Gln Ala Ile Gly Leu 965 970 975 ttt tcg ata ttc ttcatc atc ctt gcg ttt gtg atg aat tct gtg gct 2976 Phe Ser Ile Phe Phe IleIle Leu Ala Phe Val Met Asn Ser Val Ala 980 985 990 ttt att gga tcc aacctc tgg ctc agt gct tgg acc agt gac tct aaa 3024 Phe Ile Gly Ser Asn LeuTrp Leu Ser Ala Trp Thr Ser Asp Ser Lys 995 1000 1005 atc ttc aat agcacc gac tat cca gca tct cag agg gac atg aga 3069 Ile Phe Asn Ser Thr AspTyr Pro Ala Ser Gln Arg Asp Met Arg 1010 1015 1020 gtt gga gtc tac ggagct ctg gga tta gcc caa ggt ata ttt gtg 3114 Val Gly Val Tyr Gly Ala LeuGly Leu Ala Gln Gly Ile Phe Val 1025 1030 1035 ttc ata gca cat ttc tggagt gcc ttt ggt ttc gtc cat gca tca 3159 Phe Ile Ala His Phe Trp Ser AlaPhe Gly Phe Val His Ala Ser 1040 1045 1050 aat atc ttg cac aag caa ctgctg aac aat atc ctt cga gca cct 3204 Asn Ile Leu His Lys Gln Leu Leu AsnAsn Ile Leu Arg Ala Pro 1055 1060 1065 atg aga ttt ttt gac aca aca cccaca ggc cgg att gtg aac agg 3249 Met Arg Phe Phe Asp Thr Thr Pro Thr GlyArg Ile Val Asn Arg 1070 1075 1080 ttt gcc ggc gat att tcc aca gtg gatgac acc ctg cct cag tcc 3294 Phe Ala Gly Asp Ile Ser Thr Val Asp Asp ThrLeu Pro Gln Ser 1085 1090 1095 ttg cgc agc tgg att aca tgc ttc ctg gggata atc agc acc ctt 3339 Leu Arg Ser Trp Ile Thr Cys Phe Leu Gly Ile IleSer Thr Leu 1100 1105 1110 gtc atg atc tgc atg gcc act cct gtc ttc accatc atc gtc att 3384 Val Met Ile Cys Met Ala Thr Pro Val Phe Thr Ile IleVal Ile 1115 1120 1125 cct ctt ggc att att tat gta tct gtt cag atg ttttat gtg tct 3429 Pro Leu Gly Ile Ile Tyr Val Ser Val Gln Met Phe Tyr ValSer 1130 1135 1140 acc tcc cgc cag ctg agg cgt ctg gac tct gtc acc aggtcc cca 3474 Thr Ser Arg Gln Leu Arg Arg Leu Asp Ser Val Thr Arg Ser Pro1145 1150 1155 atc tac tct cac ttc agc gag acc gta tca ggt ttg cca gttatc 3519 Ile Tyr Ser His Phe Ser Glu Thr Val Ser Gly Leu Pro Val Ile1160 1165 1170 cgt gcc ttt gag cac cag cag cga ttt ctg aaa cac aat gaggag 3564 Arg Ala Phe Glu His Gln Gln Arg Phe Leu Lys His Asn Glu Glu1175 1180 1185 agg att gac acc aac cag aaa tgt gtc ttt tcc tgg atc acctcc 3609 Arg Ile Asp Thr Asn Gln Lys Cys Val Phe Ser Trp Ile Thr Ser1190 1195 1200 aac agg tgg ctt gca att cgc ctg gag ctg gtt ggg aac ctgact 3654 Asn Arg Trp Leu Ala Ile Arg Leu Glu Leu Val Gly Asn Leu Thr1205 1210 1215 gtc ttc ttt tca gcc ttg atg atg gtt att tat aga gat acccta 3699 Val Phe Phe Ser Ala Leu Met Met Val Ile Tyr Arg Asp Thr Leu1220 1225 1230 agt ggg gac act gtt ggc ttt gtt ctg tcc aat gca ctc aatatc 3744 Ser Gly Asp Thr Val Gly Phe Val Leu Ser Asn Ala Leu Asn Ile1235 1240 1245 aca caa acc ctg aac tgg ctg gtg agg atg aca tca gaa atagag 3789 Thr Gln Thr Leu Asn Trp Leu Val Arg Met Thr Ser Glu Ile Glu1250 1255 1260 acc aac att gtg gct gtt gag cga ata act gag tac aca aaagtg 3834 Thr Asn Ile Val Ala Val Glu Arg Ile Thr Glu Tyr Thr Lys Val1265 1270 1275 gaa aat gag gca ccc tgg gtg act gat aag agg cct ccg ccagat 3879 Glu Asn Glu Ala Pro Trp Val Thr Asp Lys Arg Pro Pro Pro Asp1280 1285 1290 tgg ccc agc aaa ggc aag atc cag ttt aac aac tac caa gtgcgg 3924 Trp Pro Ser Lys Gly Lys Ile Gln Phe Asn Asn Tyr Gln Val Arg1295 1300 1305 tac cga cct gag ctg gat ctg gtc ctc aga ggg atc act tgtgac 3969 Tyr Arg Pro Glu Leu Asp Leu Val Leu Arg Gly Ile Thr Cys Asp1310 1315 1320 atc ggt agc atg gag aag att ggt gtg gtg ggc agg aca ggagct 4014 Ile Gly Ser Met Glu Lys Ile Gly Val Val Gly Arg Thr Gly Ala1325 1330 1335 gga aag tca tcc ctc aca aac tgc ctc ttc aga atc tta gaggct 4059 Gly Lys Ser Ser Leu Thr Asn Cys Leu Phe Arg Ile Leu Glu Ala1340 1345 1350 gcc ggt ggt cag att atc att gat gga gta gat att gct tccatt 4104 Ala Gly Gly Gln Ile Ile Ile Asp Gly Val Asp Ile Ala Ser Ile1355 1360 1365 ggg ctc cac gac ctc cga gag aag ctg acc atc atc ccc caggac 4149 Gly Leu His Asp Leu Arg Glu Lys Leu Thr Ile Ile Pro Gln Asp1370 1375 1380 ccc atc ctg ttc tct gga agc ctg agg atg aat ctc gac cctttc 4194 Pro Ile Leu Phe Ser Gly Ser Leu Arg Met Asn Leu Asp Pro Phe1385 1390 1395 aac aac tac tca gat gag gag att tgg aag gcc ttg gag ctggct 4239 Asn Asn Tyr Ser Asp Glu Glu Ile Trp Lys Ala Leu Glu Leu Ala1400 1405 1410 cac ctc aag tct ttt gtg gcc agc ctg caa ctt ggg tta tcccac 4284 His Leu Lys Ser Phe Val Ala Ser Leu Gln Leu Gly Leu Ser His1415 1420 1425 gaa gtt aca gag gct ggt ggc aac ctg agc ata ggc cag aggcag 4329 Glu Val Thr Glu Ala Gly Gly Asn Leu Ser Ile Gly Gln Arg Gln1430 1435 1440 ctg ctg tgc ctg ggc agg gct ctg ctt cgg aaa tcc aag atcctg 4374 Leu Leu Cys Leu Gly Arg Ala Leu Leu Arg Lys Ser Lys Ile Leu1445 1450 1455 gtc ctg gat gag gcc act gct gcg gtg gat cta gag aca gacaac 4419 Val Leu Asp Glu Ala Thr Ala Ala Val Asp Leu Glu Thr Asp Asn1460 1465 1470 ctc att cag acg acc atc caa aac gag ttc gcc cac tgc acagtg 4464 Leu Ile Gln Thr Thr Ile Gln Asn Glu Phe Ala His Cys Thr Val1475 1480 1485 atc acc atc gcc cac agg ctg cat acc atc atg gac agt gacaag 4509 Ile Thr Ile Ala His Arg Leu His Thr Ile Met Asp Ser Asp Lys1490 1495 1500 gta atg gtc cta gac aac ggg aag att ata gag tac ggc agccct 4554 Val Met Val Leu Asp Asn Gly Lys Ile Ile Glu Tyr Gly Ser Pro1505 1510 1515 gaa gaa ctg cta caa atc cct gga ccc ttt tac ttt atg gctaag 4599 Glu Glu Leu Leu Gln Ile Pro Gly Pro Phe Tyr Phe Met Ala Lys1520 1525 1530 gaa gct ggc att gag aat gtg aac agc gca ccg gtc gcc 4638Glu Ala Gly Ile Glu Asn Val Asn Ser Ala Pro Val 1535 1540 1545 6 1545PRT Homo sapiens 6 Met Leu Glu Lys Phe Cys Asn Ser Thr Phe Trp Asn SerSer Phe Leu 1 5 10 15 Asp Ser Pro Glu Ala Asp Leu Pro Leu Cys Phe GluGln Thr Val Leu 20 25 30 Val Trp Ile Pro Leu Gly Phe Leu Trp Leu Leu AlaPro Trp Gln Leu 35 40 45 Leu His Val Tyr Lys Ser Arg Thr Lys Arg Ser SerThr Thr Lys Leu 50 55 60 Tyr Leu Ala Lys Gln Val Phe Val Gly Phe Leu LeuIle Leu Ala Ala 65 70 75 80 Ile Glu Leu Ala Leu Val Leu Thr Glu Asp SerGly Gln Ala Thr Val 85 90 95 Pro Ala Val Arg Tyr Thr Asn Pro Ser Leu TyrLeu Gly Thr Trp Leu 100 105 110 Leu Val Leu Leu Ile Gln Tyr Ser Arg GlnTrp Cys Val Gln Lys Asn 115 120 125 Ser Trp Phe Leu Ser Leu Phe Trp IleLeu Ser Ile Leu Cys Gly Thr 130 135 140 Phe Gln Phe Gln Thr Leu Ile ArgThr Leu Leu Gln Gly Asp Asn Ser 145 150 155 160 Asn Leu Ala Tyr Ser CysLeu Phe Phe Ile Ser Tyr Gly Phe Gln Ile 165 170 175 Leu Ile Leu Ile PheSer Ala Phe Ser Glu Asn Asn Glu Ser Ser Asn 180 185 190 Asn Pro Ser SerIle Ala Ser Phe Leu Ser Ser Ile Thr Tyr Ser Trp 195 200 205 Tyr Asp SerIle Ile Leu Lys Gly Tyr Lys Arg Pro Leu Thr Leu Glu 210 215 220 Asp ValTrp Glu Val Asp Glu Glu Met Lys Thr Lys Thr Leu Val Ser 225 230 235 240Lys Phe Glu Thr His Met Lys Arg Glu Leu Gln Lys Ala Arg Arg Ala 245 250255 Leu Gln Arg Arg Gln Glu Lys Ser Ser Gln Gln Asn Ser Gly Ala Arg 260265 270 Leu Pro Gly Leu Asn Lys Asn Gln Ser Gln Ser Gln Asp Ala Leu Val275 280 285 Leu Glu Asp Val Glu Lys Lys Lys Lys Lys Ser Gly Thr Lys LysAsp 290 295 300 Val Pro Lys Ser Trp Leu Met Lys Ala Leu Phe Lys Thr PheTyr Met 305 310 315 320 Val Leu Leu Lys Ser Phe Leu Leu Lys Leu Val AsnAsp Ile Phe Thr 325 330 335 Phe Val Ser Pro Gln Leu Leu Lys Leu Leu IleSer Phe Ala Ser Asp 340 345 350 Arg Asp Thr Tyr Leu Trp Ile Gly Tyr LeuCys Ala Ile Leu Leu Phe 355 360 365 Thr Ala Ala Leu Ile Gln Ser Phe CysLeu Gln Cys Tyr Phe Gln Leu 370 375 380 Cys Phe Lys Leu Gly Val Lys ValArg Thr Ala Ile Met Ala Ser Val 385 390 395 400 Tyr Lys Lys Ala Leu ThrLeu Ser Asn Leu Ala Arg Lys Glu Tyr Thr 405 410 415 Val Gly Glu Thr ValAsn Leu Met Ser Val Asp Ala Gln Lys Leu Met 420 425 430 Asp Val Thr AsnPhe Met His Met Leu Trp Ser Ser Val Leu Gln Ile 435 440 445 Val Leu SerIle Phe Phe Leu Trp Arg Glu Leu Gly Pro Ser Val Leu 450 455 460 Ala GlyVal Gly Val Met Val Leu Val Ile Pro Ile Asn Ala Ile Leu 465 470 475 480Ser Thr Lys Ser Lys Thr Ile Gln Val Lys Asn Met Lys Asn Lys Asp 485 490495 Lys Arg Leu Lys Ile Met Asn Glu Ile Leu Ser Gly Ile Lys Ile Leu 500505 510 Lys Tyr Phe Ala Trp Glu Pro Ser Phe Arg Asp Gln Val Gln Asn Leu515 520 525 Arg Lys Lys Glu Leu Lys Asn Leu Leu Ala Phe Ser Gln Leu GlnCys 530 535 540 Val Val Ile Phe Val Phe Gln Leu Thr Pro Val Leu Val SerVal Val 545 550 555 560 Thr Phe Ser Val Tyr Val Leu Val Asp Ser Asn AsnIle Leu Asp Ala 565 570 575 Gln Lys Ala Phe Thr Ser Ile Thr Leu Phe AsnIle Leu Arg Phe Pro 580 585 590 Leu Ser Met Leu Pro Met Met Ile Ser SerMet Leu Gln Ala Ser Val 595 600 605 Ser Thr Glu Arg Leu Glu Lys Tyr LeuGly Gly Asp Asp Leu Asp Thr 610 615 620 Ser Ala Ile Arg His Asp Cys AsnPhe Asp Lys Ala Met Gln Phe Ser 625 630 635 640 Glu Ala Ser Phe Thr TrpGlu His Asp Ser Glu Ala Thr Val Arg Asp 645 650 655 Val Asn Leu Asp IleMet Ala Gly Gln Leu Val Ala Val Ile Gly Pro 660 665 670 Val Gly Ser GlyLys Ser Ser Leu Ile Ser Ala Met Leu Gly Glu Met 675 680 685 Glu Asn ValHis Gly His Ile Thr Ile Lys Gly Thr Thr Ala Tyr Val 690 695 700 Pro GlnGln Ser Trp Ile Gln Asn Gly Thr Ile Lys Asp Asn Ile Leu 705 710 715 720Phe Gly Thr Glu Phe Asn Glu Lys Arg Tyr Gln Gln Val Leu Glu Ala 725 730735 Cys Ala Leu Leu Pro Asp Leu Glu Met Leu Pro Gly Gly Asp Leu Ala 740745 750 Glu Ile Gly Glu Lys Gly Ile Asn Leu Ser Gly Gly Gln Lys Gln Arg755 760 765 Ile Ser Leu Ala Arg Ala Thr Tyr Gln Asn Leu Asp Ile Tyr LeuLeu 770 775 780 Asp Asp Pro Leu Ser Ala Val Asp Ala His Val Gly Lys HisIle Phe 785 790 795 800 Asn Lys Val Leu Gly Pro Asn Gly Leu Leu Lys GlyLys Thr Arg Leu 805 810 815 Leu Val Thr His Ser Met His Phe Leu Pro GlnVal Asp Glu Ile Val 820 825 830 Val Leu Gly Asn Gly Thr Ile Val Glu LysGly Ser Tyr Ser Ala Leu 835 840 845 Leu Ala Lys Lys Gly Glu Phe Ala LysAsn Leu Lys Thr Phe Leu Arg 850 855 860 His Thr Gly Pro Glu Glu Glu AlaThr Val His Asp Gly Ser Glu Glu 865 870 875 880 Glu Asp Asp Asp Tyr GlyLeu Ile Ser Ser Val Glu Glu Ile Pro Glu 885 890 895 Asp Ala Ala Ser IleThr Met Arg Arg Glu Asn Ser Phe Arg Arg Thr 900 905 910 Leu Ser Arg SerSer Arg Ser Asn Gly Arg His Leu Lys Ser Leu Arg 915 920 925 Asn Ser LeuLys Thr Arg Asn Val Asn Ser Leu Lys Glu Asp Glu Glu 930 935 940 Leu ValLys Gly Gln Lys Leu Ile Lys Lys Glu Phe Ile Glu Thr Gly 945 950 955 960Lys Val Lys Phe Ser Ile Tyr Leu Glu Tyr Leu Gln Ala Ile Gly Leu 965 970975 Phe Ser Ile Phe Phe Ile Ile Leu Ala Phe Val Met Asn Ser Val Ala 980985 990 Phe Ile Gly Ser Asn Leu Trp Leu Ser Ala Trp Thr Ser Asp Ser Lys995 1000 1005 Ile Phe Asn Ser Thr Asp Tyr Pro Ala Ser Gln Arg Asp MetArg 1010 1015 1020 Val Gly Val Tyr Gly Ala Leu Gly Leu Ala Gln Gly IlePhe Val 1025 1030 1035 Phe Ile Ala His Phe Trp Ser Ala Phe Gly Phe ValHis Ala Ser 1040 1045 1050 Asn Ile Leu His Lys Gln Leu Leu Asn Asn IleLeu Arg Ala Pro 1055 1060 1065 Met Arg Phe Phe Asp Thr Thr Pro Thr GlyArg Ile Val Asn Arg 1070 1075 1080 Phe Ala Gly Asp Ile Ser Thr Val AspAsp Thr Leu Pro Gln Ser 1085 1090 1095 Leu Arg Ser Trp Ile Thr Cys PheLeu Gly Ile Ile Ser Thr Leu 1100 1105 1110 Val Met Ile Cys Met Ala ThrPro Val Phe Thr Ile Ile Val Ile 1115 1120 1125 Pro Leu Gly Ile Ile TyrVal Ser Val Gln Met Phe Tyr Val Ser 1130 1135 1140 Thr Ser Arg Gln LeuArg Arg Leu Asp Ser Val Thr Arg Ser Pro 1145 1150 1155 Ile Tyr Ser HisPhe Ser Glu Thr Val Ser Gly Leu Pro Val Ile 1160 1165 1170 Arg Ala PheGlu His Gln Gln Arg Phe Leu Lys His Asn Glu Glu 1175 1180 1185 Arg IleAsp Thr Asn Gln Lys Cys Val Phe Ser Trp Ile Thr Ser 1190 1195 1200 AsnArg Trp Leu Ala Ile Arg Leu Glu Leu Val Gly Asn Leu Thr 1205 1210 1215Val Phe Phe Ser Ala Leu Met Met Val Ile Tyr Arg Asp Thr Leu 1220 12251230 Ser Gly Asp Thr Val Gly Phe Val Leu Ser Asn Ala Leu Asn Ile 12351240 1245 Thr Gln Thr Leu Asn Trp Leu Val Arg Met Thr Ser Glu Ile Glu1250 1255 1260 Thr Asn Ile Val Ala Val Glu Arg Ile Thr Glu Tyr Thr LysVal 1265 1270 1275 Glu Asn Glu Ala Pro Trp Val Thr Asp Lys Arg Pro ProPro Asp 1280 1285 1290 Trp Pro Ser Lys Gly Lys Ile Gln Phe Asn Asn TyrGln Val Arg 1295 1300 1305 Tyr Arg Pro Glu Leu Asp Leu Val Leu Arg GlyIle Thr Cys Asp 1310 1315 1320 Ile Gly Ser Met Glu Lys Ile Gly Val ValGly Arg Thr Gly Ala 1325 1330 1335 Gly Lys Ser Ser Leu Thr Asn Cys LeuPhe Arg Ile Leu Glu Ala 1340 1345 1350 Ala Gly Gly Gln Ile Ile Ile AspGly Val Asp Ile Ala Ser Ile 1355 1360 1365 Gly Leu His Asp Leu Arg GluLys Leu Thr Ile Ile Pro Gln Asp 1370 1375 1380 Pro Ile Leu Phe Ser GlySer Leu Arg Met Asn Leu Asp Pro Phe 1385 1390 1395 Asn Asn Tyr Ser AspGlu Glu Ile Trp Lys Ala Leu Glu Leu Ala 1400 1405 1410 His Leu Lys SerPhe Val Ala Ser Leu Gln Leu Gly Leu Ser His 1415 1420 1425 Glu Val ThrGlu Ala Gly Gly Asn Leu Ser Ile Gly Gln Arg Gln 1430 1435 1440 Leu LeuCys Leu Gly Arg Ala Leu Leu Arg Lys Ser Lys Ile Leu 1445 1450 1455 ValLeu Asp Glu Ala Thr Ala Ala Val Asp Leu Glu Thr Asp Asn 1460 1465 1470Leu Ile Gln Thr Thr Ile Gln Asn Glu Phe Ala His Cys Thr Val 1475 14801485 Ile Thr Ile Ala His Arg Leu His Thr Ile Met Asp Ser Asp Lys 14901495 1500 Val Met Val Leu Asp Asn Gly Lys Ile Ile Glu Tyr Gly Ser Pro1505 1510 1515 Glu Glu Leu Leu Gln Ile Pro Gly Pro Phe Tyr Phe Met AlaLys 1520 1525 1530 Glu Ala Gly Ile Glu Asn Val Asn Ser Ala Pro Val 15351540 1545 7 4637 DNA Homo sapiens CDS (1)..(4635) 7 atg ctg gag aag ttctgc aac tct act ttt tgg aat tcc tca ttc ctg 48 Met Leu Glu Lys Phe CysAsn Ser Thr Phe Trp Asn Ser Ser Phe Leu 1 5 10 15 gac agt ccg gag gcagac ctg cca ctt tgt ttt gag caa act gtt ctg 96 Asp Ser Pro Glu Ala AspLeu Pro Leu Cys Phe Glu Gln Thr Val Leu 20 25 30 gtg tgg att ccc ttg ggcttc cta tgg ctc ctg gcc ccc tgg cag ctt 144 Val Trp Ile Pro Leu Gly PheLeu Trp Leu Leu Ala Pro Trp Gln Leu 35 40 45 ctc cac gtg tat aaa tcc aggacc aag aga tcc tct acc acc aaa ctc 192 Leu His Val Tyr Lys Ser Arg ThrLys Arg Ser Ser Thr Thr Lys Leu 50 55 60 tat ctt gct aag cag gta ttc gttggt ttt ctt ctt att cta gca gcc 240 Tyr Leu Ala Lys Gln Val Phe Val GlyPhe Leu Leu Ile Leu Ala Ala 65 70 75 80 ata gag ctg gcc ctt gta ctc acagaa gac tct gga caa gcc aca gtc 288 Ile Glu Leu Ala Leu Val Leu Thr GluAsp Ser Gly Gln Ala Thr Val 85 90 95 cct gct gtt cga tat acc aat cca agcctc tac cta ggc aca tgg ctc 336 Pro Ala Val Arg Tyr Thr Asn Pro Ser LeuTyr Leu Gly Thr Trp Leu 100 105 110 ctg gtt ttg ctg atc caa tac agc agacaa tgg tgt gta cag aaa aac 384 Leu Val Leu Leu Ile Gln Tyr Ser Arg GlnTrp Cys Val Gln Lys Asn 115 120 125 tcc tgg ttc ctg tcc cta ttc tgg attctc tcg ata ctc tgt ggc act 432 Ser Trp Phe Leu Ser Leu Phe Trp Ile LeuSer Ile Leu Cys Gly Thr 130 135 140 ttc caa ttt cag act ctg atc cgg acactc tta cag ggt gac aat tct 480 Phe Gln Phe Gln Thr Leu Ile Arg Thr LeuLeu Gln Gly Asp Asn Ser 145 150 155 160 aat cta gcc tac tcc tgc ctg ttcttc atc tcc tac gga ttc cag atc 528 Asn Leu Ala Tyr Ser Cys Leu Phe PheIle Ser Tyr Gly Phe Gln Ile 165 170 175 ctg atc ctg atc ttt tca gca ttttca gaa aat aat gag tca tca aat 576 Leu Ile Leu Ile Phe Ser Ala Phe SerGlu Asn Asn Glu Ser Ser Asn 180 185 190 aat cca tca tcc ata gct tca ttcctg agt agc att acc tac agc tgg 624 Asn Pro Ser Ser Ile Ala Ser Phe LeuSer Ser Ile Thr Tyr Ser Trp 195 200 205 tat gac agc atc att ctg aaa ggctac aag cgt cct ctg aca ctc gag 672 Tyr Asp Ser Ile Ile Leu Lys Gly TyrLys Arg Pro Leu Thr Leu Glu 210 215 220 gat gtc tgg gaa gtt gat gaa gagatg aaa acc aag aca tta gtg agc 720 Asp Val Trp Glu Val Asp Glu Glu MetLys Thr Lys Thr Leu Val Ser 225 230 235 240 aag ttt gaa acg cac atg aagaga gag ctg cag aaa gcc agg cgg gca 768 Lys Phe Glu Thr His Met Lys ArgGlu Leu Gln Lys Ala Arg Arg Ala 245 250 255 ctc cag aga cgg cag gag aagagc tcc cag cag aac tct gga gcc agg 816 Leu Gln Arg Arg Gln Glu Lys SerSer Gln Gln Asn Ser Gly Ala Arg 260 265 270 ctg cct ggc ttg aac aag aatcag agt caa agc caa gat gcc ctt gtc 864 Leu Pro Gly Leu Asn Lys Asn GlnSer Gln Ser Gln Asp Ala Leu Val 275 280 285 ctg gaa gat gtt gaa aag aaaaaa aag aag tct ggg acc aaa aaa gat 912 Leu Glu Asp Val Glu Lys Lys LysLys Lys Ser Gly Thr Lys Lys Asp 290 295 300 gtt cca aaa tcc tgg ttg atgaag gct ctg ttc aaa act ttc tac atg 960 Val Pro Lys Ser Trp Leu Met LysAla Leu Phe Lys Thr Phe Tyr Met 305 310 315 320 gtg ctc ctg aaa tca ttccta ctg aag cta gtg aat gac atc ttc acg 1008 Val Leu Leu Lys Ser Phe LeuLeu Lys Leu Val Asn Asp Ile Phe Thr 325 330 335 ttt gtg agt cct cag ctgctg aaa ttg ctg atc tcc ttt gca agt gac 1056 Phe Val Ser Pro Gln Leu LeuLys Leu Leu Ile Ser Phe Ala Ser Asp 340 345 350 cgt gac aca tat ttg tggatt gga tat ctc tgt gca atc ctc tta ttc 1104 Arg Asp Thr Tyr Leu Trp IleGly Tyr Leu Cys Ala Ile Leu Leu Phe 355 360 365 act gcg gct ctc att cagtct ttc tgc ctt cag tgt tat ttc caa ctg 1152 Thr Ala Ala Leu Ile Gln SerPhe Cys Leu Gln Cys Tyr Phe Gln Leu 370 375 380 tgc ttc aag ctg ggt gtaaaa gta cgg aca gct atc atg gct tct gta 1200 Cys Phe Lys Leu Gly Val LysVal Arg Thr Ala Ile Met Ala Ser Val 385 390 395 400 tat aag aag gca ttgacc cta tcc aac ttg gcc agg aag gag tac acc 1248 Tyr Lys Lys Ala Leu ThrLeu Ser Asn Leu Ala Arg Lys Glu Tyr Thr 405 410 415 gtt gga gaa aca gtgaac ctg atg tct gtg gat gcc cag aag ctc atg 1296 Val Gly Glu Thr Val AsnLeu Met Ser Val Asp Ala Gln Lys Leu Met 420 425 430 gat gtg acc aac ttcatg cac atg ctg tgg tca agt gtt cta cag att 1344 Asp Val Thr Asn Phe MetHis Met Leu Trp Ser Ser Val Leu Gln Ile 435 440 445 gtc tta tct atc ttcttc cta tgg aga gag ttg gga ccc tca gtc tta 1392 Val Leu Ser Ile Phe PheLeu Trp Arg Glu Leu Gly Pro Ser Val Leu 450 455 460 gca ggt gtt ggg gtgatg gtg ctt gta atc cca att aat gcg ata ctg 1440 Ala Gly Val Gly Val MetVal Leu Val Ile Pro Ile Asn Ala Ile Leu 465 470 475 480 tcc acc aag agtaag acc att cag gtc aaa aat atg aag aat aaa gac 1488 Ser Thr Lys Ser LysThr Ile Gln Val Lys Asn Met Lys Asn Lys Asp 485 490 495 aaa cgt tta aagatc atg aat gag att ctt agt gga atc aag atc ctg 1536 Lys Arg Leu Lys IleMet Asn Glu Ile Leu Ser Gly Ile Lys Ile Leu 500 505 510 aaa tat ttt gcctgg gaa cct tca ttc aga gac caa gta caa aac ctc 1584 Lys Tyr Phe Ala TrpGlu Pro Ser Phe Arg Asp Gln Val Gln Asn Leu 515 520 525 cgg aag aaa gagctc aag aac ctg ctg gcc ttt agt caa cta cag tgt 1632 Arg Lys Lys Glu LeuLys Asn Leu Leu Ala Phe Ser Gln Leu Gln Cys 530 535 540 gta gta ata ttcgtc ttc cag tta act cca gtc ctg gta tct gtg gtc 1680 Val Val Ile Phe ValPhe Gln Leu Thr Pro Val Leu Val Ser Val Val 545 550 555 560 aca ttt tctgtt tat gtc ctg gtg gat agc aac aat att ttg gat gca 1728 Thr Phe Ser ValTyr Val Leu Val Asp Ser Asn Asn Ile Leu Asp Ala 565 570 575 caa aag gccttc acc tcc att acc ctc ttc aat atc ctg cgc ttt ccc 1776 Gln Lys Ala PheThr Ser Ile Thr Leu Phe Asn Ile Leu Arg Phe Pro 580 585 590 ctg agc atgctt ccc atg atg atc tcc tcc atg ctc cag gcc agt gtt 1824 Leu Ser Met LeuPro Met Met Ile Ser Ser Met Leu Gln Ala Ser Val 595 600 605 tcc aca gagcgg cta gag aag tac ttg gga ggg gat gac ttg gac aca 1872 Ser Thr Glu ArgLeu Glu Lys Tyr Leu Gly Gly Asp Asp Leu Asp Thr 610 615 620 tct gcc attcga cat gac tgc aat ttt gac aaa gcc atg cag ttt tct 1920 Ser Ala Ile ArgHis Asp Cys Asn Phe Asp Lys Ala Met Gln Phe Ser 625 630 635 640 gag gcctcc ttt acc tgg gaa cat gat tcg gaa gcc aca gtc cga gat 1968 Glu Ala SerPhe Thr Trp Glu His Asp Ser Glu Ala Thr Val Arg Asp 645 650 655 gtg aacctg gac att atg gca ggc caa ctt gtg gct gtg ata ggc cct 2016 Val Asn LeuAsp Ile Met Ala Gly Gln Leu Val Ala Val Ile Gly Pro 660 665 670 gtc ggctct ggg aaa tcc tcc ttg ata tca gcc atg ctg gga gaa atg 2064 Val Gly SerGly Lys Ser Ser Leu Ile Ser Ala Met Leu Gly Glu Met 675 680 685 gaa aatgtc cac ggg cac atc acc atc aag ggc acc act gcc tat gtc 2112 Glu Asn ValHis Gly His Ile Thr Ile Lys Gly Thr Thr Ala Tyr Val 690 695 700 cca cagcag tcc tgg att cag aat ggc acc ata aag gac aac atc ctt 2160 Pro Gln GlnSer Trp Ile Gln Asn Gly Thr Ile Lys Asp Asn Ile Leu 705 710 715 720 tttgga aca gag ttt aat gaa aag agg tac cag caa gta ctg gag gcc 2208 Phe GlyThr Glu Phe Asn Glu Lys Arg Tyr Gln Gln Val Leu Glu Ala 725 730 735 tgtgct ctc ctc cca gac ttg gaa atg ctg cct gga gga gat ttg gct 2256 Cys AlaLeu Leu Pro Asp Leu Glu Met Leu Pro Gly Gly Asp Leu Ala 740 745 750 gagatt gga gag aag ggt ata aat ctt agt ggg ggt cag aag cag cgg 2304 Glu IleGly Glu Lys Gly Ile Asn Leu Ser Gly Gly Gln Lys Gln Arg 755 760 765 atcagc ctg gcc aga gct acc tac caa aat tta gac atc tat ctt cta 2352 Ile SerLeu Ala Arg Ala Thr Tyr Gln Asn Leu Asp Ile Tyr Leu Leu 770 775 780 gatgac ccc ctg tct gca gtg gat gct cat gta gga aaa cat att ttt 2400 Asp AspPro Leu Ser Ala Val Asp Ala His Val Gly Lys His Ile Phe 785 790 795 800aat aag gtc ttg ggc ccc aat ggc ctg ttg aaa ggc aag act cga ctc 2448 AsnLys Val Leu Gly Pro Asn Gly Leu Leu Lys Gly Lys Thr Arg Leu 805 810 815ttg gtt aca cat agc atg cac ttt ctt cct caa gtg gat gag att gta 2496 LeuVal Thr His Ser Met His Phe Leu Pro Gln Val Asp Glu Ile Val 820 825 830gtt ctg ggg aat gga aca att gta gag aaa gga tcc tac agt gct ctc 2544 ValLeu Gly Asn Gly Thr Ile Val Glu Lys Gly Ser Tyr Ser Ala Leu 835 840 845ctg gcc aaa aaa gga gag ttt gct aag aat ctg aag aca ttt cta aga 2592 LeuAla Lys Lys Gly Glu Phe Ala Lys Asn Leu Lys Thr Phe Leu Arg 850 855 860cat aca ggc cct gaa gag gaa gcc aca gtc cat gat ggc agt gaa gaa 2640 HisThr Gly Pro Glu Glu Glu Ala Thr Val His Asp Gly Ser Glu Glu 865 870 875880 gaa gac gat gac tat ggg ctg ata tcc agt gtg gaa gag atc ccc gaa 2688Glu Asp Asp Asp Tyr Gly Leu Ile Ser Ser Val Glu Glu Ile Pro Glu 885 890895 gat gca gcc tcc ata acc atg aga aga gag aac agc ttt cgt cga aca 2736Asp Ala Ala Ser Ile Thr Met Arg Arg Glu Asn Ser Phe Arg Arg Thr 900 905910 ctt agc cgc agt tct agg tcc aat ggc agg cat ctg aag tcc ctg aga 2784Leu Ser Arg Ser Ser Arg Ser Asn Gly Arg His Leu Lys Ser Leu Arg 915 920925 aac tcc ttg aaa act cgg aat gtg aat agc ctg aag gaa gac gaa gaa 2832Asn Ser Leu Lys Thr Arg Asn Val Asn Ser Leu Lys Glu Asp Glu Glu 930 935940 cta gtg aaa gga caa aaa cta att aag aag gaa ttc ata gaa act gga 2880Leu Val Lys Gly Gln Lys Leu Ile Lys Lys Glu Phe Ile Glu Thr Gly 945 950955 960 aag gtg aag ttc tcc atc tac ctg gag tac cta caa gca ata gga ttg2928 Lys Val Lys Phe Ser Ile Tyr Leu Glu Tyr Leu Gln Ala Ile Gly Leu 965970 975 ttt tcg ata ttc ttc atc atc ctt gcg ttt gtg atg aat tct gtg gct2976 Phe Ser Ile Phe Phe Ile Ile Leu Ala Phe Val Met Asn Ser Val Ala 980985 990 ttt att gga tcc aac ctc tgg ctc agt gct tgg acc agt gac tct aaa3024 Phe Ile Gly Ser Asn Leu Trp Leu Ser Ala Trp Thr Ser Asp Ser Lys 9951000 1005 atc ttc aat agc acc gac tat cca gca tct cag agg gac atg aga3069 Ile Phe Asn Ser Thr Asp Tyr Pro Ala Ser Gln Arg Asp Met Arg 10101015 1020 gtt gga gtc tac gga gct ctg gga tta gcc caa ggt ata ttt gtg3114 Val Gly Val Tyr Gly Ala Leu Gly Leu Ala Gln Gly Ile Phe Val 10251030 1035 ttc ata gca cat ttc tgg agt gcc ttt ggt ttc gtc cat gca tca3159 Phe Ile Ala His Phe Trp Ser Ala Phe Gly Phe Val His Ala Ser 10401045 1050 aat atc ttg cac aag caa ctg ctg aac aat atc ctt cga gca cct3204 Asn Ile Leu His Lys Gln Leu Leu Asn Asn Ile Leu Arg Ala Pro 10551060 1065 atg aga ttt ttt gac aca aca ccc aca ggc cgg att gtg aac agg3249 Met Arg Phe Phe Asp Thr Thr Pro Thr Gly Arg Ile Val Asn Arg 10701075 1080 ttt gcc ggc gat att tcc aca gtg gat gac acc ctg cct cag tcc3294 Phe Ala Gly Asp Ile Ser Thr Val Asp Asp Thr Leu Pro Gln Ser 10851090 1095 ttg cgc agc tgg att aca tgc ttc ctg ggg ata atc agc acc ctt3339 Leu Arg Ser Trp Ile Thr Cys Phe Leu Gly Ile Ile Ser Thr Leu 11001105 1110 gtc atg atc tgc atg gcc act cct gtc ttc acc atc atc gtc att3384 Val Met Ile Cys Met Ala Thr Pro Val Phe Thr Ile Ile Val Ile 11151120 1125 cct ctt ggc att att tat gta tct gtt cag atg ttt tat gtg tct3429 Pro Leu Gly Ile Ile Tyr Val Ser Val Gln Met Phe Tyr Val Ser 11301135 1140 acc tcc cgc cag ctg agg cgt ctg gac tct gtc acc agg tcc cca3474 Thr Ser Arg Gln Leu Arg Arg Leu Asp Ser Val Thr Arg Ser Pro 11451150 1155 atc tac tct cac ttc agc gag acc gta tca ggt ttg cca gtt atc3519 Ile Tyr Ser His Phe Ser Glu Thr Val Ser Gly Leu Pro Val Ile 11601165 1170 cgt gcc ttt gag cac cag cag cga ttt ctg aaa cac aat gag gag3564 Arg Ala Phe Glu His Gln Gln Arg Phe Leu Lys His Asn Glu Glu 11751180 1185 agg att gac acc aac cag aaa tgt gtc ttt tcc tgg atc acc tcc3609 Arg Ile Asp Thr Asn Gln Lys Cys Val Phe Ser Trp Ile Thr Ser 11901195 1200 aac agg tgg ctt gca att cgc ctg gag ctg gtt ggg aac ctg act3654 Asn Arg Trp Leu Ala Ile Arg Leu Glu Leu Val Gly Asn Leu Thr 12051210 1215 gtc ttc ttt tca gcc ttg atg atg gtt att tat aga gat acc cta3699 Val Phe Phe Ser Ala Leu Met Met Val Ile Tyr Arg Asp Thr Leu 12201225 1230 agt ggg gac act gtt ggc ttt gtt ctg tcc aat gca ctc aat atc3744 Ser Gly Asp Thr Val Gly Phe Val Leu Ser Asn Ala Leu Asn Ile 12351240 1245 aca caa acc ctg aac tgg ctg gtg agg atg aca tca gaa ata gag3789 Thr Gln Thr Leu Asn Trp Leu Val Arg Met Thr Ser Glu Ile Glu 12501255 1260 acc aac att gtg gct gtt gag cga ata act gag tac aca aaa gtg3834 Thr Asn Ile Val Ala Val Glu Arg Ile Thr Glu Tyr Thr Lys Val 12651270 1275 gaa aat gag gca ccc tgg gtg act gat aag agg cct ccg cca gat3879 Glu Asn Glu Ala Pro Trp Val Thr Asp Lys Arg Pro Pro Pro Asp 12801285 1290 tgg ccc agc aaa ggc aag atc cag ttt aac aac tac caa gtg cgg3924 Trp Pro Ser Lys Gly Lys Ile Gln Phe Asn Asn Tyr Gln Val Arg 12951300 1305 tac cga cct gag ctg gat ctg gtc ctc aga ggg atc act tgt gac3969 Tyr Arg Pro Glu Leu Asp Leu Val Leu Arg Gly Ile Thr Cys Asp 13101315 1320 atc ggt agc atg gag aag att ggt gtg gtg ggc agg aca gga gct4014 Ile Gly Ser Met Glu Lys Ile Gly Val Val Gly Arg Thr Gly Ala 13251330 1335 gga aag tca tcc ctc aca aac tgc ctc ttc aga atc tta gag gct4059 Gly Lys Ser Ser Leu Thr Asn Cys Leu Phe Arg Ile Leu Glu Ala 13401345 1350 gcc ggt ggt cag att atc att gat gga gta gat att gct tcc att4104 Ala Gly Gly Gln Ile Ile Ile Asp Gly Val Asp Ile Ala Ser Ile 13551360 1365 ggg ctc cac gac ctc cga gag aag ctg acc atc atc ccc cag gac4149 Gly Leu His Asp Leu Arg Glu Lys Leu Thr Ile Ile Pro Gln Asp 13701375 1380 ccc atc ctg ttc tct gga agc ctg agg atg aat ctc gac cct ttc4194 Pro Ile Leu Phe Ser Gly Ser Leu Arg Met Asn Leu Asp Pro Phe 13851390 1395 aac aac tac tca gat gag gag att tgg aag gcc ttg gag ctg gct4239 Asn Asn Tyr Ser Asp Glu Glu Ile Trp Lys Ala Leu Glu Leu Ala 14001405 1410 cac ctc aag tct ttt gtg gcc agc ctg caa ctt ggg tta tcc cac4284 His Leu Lys Ser Phe Val Ala Ser Leu Gln Leu Gly Leu Ser His 14151420 1425 gaa gtt aca gag gct ggt ggc aac ctg agc ata ggc cag agg cag4329 Glu Val Thr Glu Ala Gly Gly Asn Leu Ser Ile Gly Gln Arg Gln 14301435 1440 ctg ctg tgc ctg ggc agg gct ctg ctt cgg aaa tcc aag atc ctg4374 Leu Leu Cys Leu Gly Arg Ala Leu Leu Arg Lys Ser Lys Ile Leu 14451450 1455 gtc ctg gat gag gcc act gct gcg gtg gat cta gag aca gac aac4419 Val Leu Asp Glu Ala Thr Ala Ala Val Asp Leu Glu Thr Asp Asn 14601465 1470 ctc att cag acg acc atc caa aac gag ttc gcc cac tgc aca gtg4464 Leu Ile Gln Thr Thr Ile Gln Asn Glu Phe Ala His Cys Thr Val 14751480 1485 atc acc atc gcc cac agg ctg cat acc atc atg gac agt gac aag4509 Ile Thr Ile Ala His Arg Leu His Thr Ile Met Asp Ser Asp Lys 14901495 1500 gta atg gtc cta gac aac ggg aag att ata gag tac ggc agc cct4554 Val Met Val Leu Asp Asn Gly Lys Ile Ile Glu Tyr Gly Ser Pro 15051510 1515 gaa gaa ctg cta caa atc cct gga ccc ttt tac ttt atg gct aag4599 Glu Glu Leu Leu Gln Ile Pro Gly Pro Phe Tyr Phe Met Ala Lys 15201525 1530 gaa gct ggc att gag aat gtg aac gcc aca aaa ttc gc 4637 GluAla Gly Ile Glu Asn Val Asn Ala Thr Lys Phe 1535 1540 1545 8 1545 PRTHomo sapiens 8 Met Leu Glu Lys Phe Cys Asn Ser Thr Phe Trp Asn Ser SerPhe Leu 1 5 10 15 Asp Ser Pro Glu Ala Asp Leu Pro Leu Cys Phe Glu GlnThr Val Leu 20 25 30 Val Trp Ile Pro Leu Gly Phe Leu Trp Leu Leu Ala ProTrp Gln Leu 35 40 45 Leu His Val Tyr Lys Ser Arg Thr Lys Arg Ser Ser ThrThr Lys Leu 50 55 60 Tyr Leu Ala Lys Gln Val Phe Val Gly Phe Leu Leu IleLeu Ala Ala 65 70 75 80 Ile Glu Leu Ala Leu Val Leu Thr Glu Asp Ser GlyGln Ala Thr Val 85 90 95 Pro Ala Val Arg Tyr Thr Asn Pro Ser Leu Tyr LeuGly Thr Trp Leu 100 105 110 Leu Val Leu Leu Ile Gln Tyr Ser Arg Gln TrpCys Val Gln Lys Asn 115 120 125 Ser Trp Phe Leu Ser Leu Phe Trp Ile LeuSer Ile Leu Cys Gly Thr 130 135 140 Phe Gln Phe Gln Thr Leu Ile Arg ThrLeu Leu Gln Gly Asp Asn Ser 145 150 155 160 Asn Leu Ala Tyr Ser Cys LeuPhe Phe Ile Ser Tyr Gly Phe Gln Ile 165 170 175 Leu Ile Leu Ile Phe SerAla Phe Ser Glu Asn Asn Glu Ser Ser Asn 180 185 190 Asn Pro Ser Ser IleAla Ser Phe Leu Ser Ser Ile Thr Tyr Ser Trp 195 200 205 Tyr Asp Ser IleIle Leu Lys Gly Tyr Lys Arg Pro Leu Thr Leu Glu 210 215 220 Asp Val TrpGlu Val Asp Glu Glu Met Lys Thr Lys Thr Leu Val Ser 225 230 235 240 LysPhe Glu Thr His Met Lys Arg Glu Leu Gln Lys Ala Arg Arg Ala 245 250 255Leu Gln Arg Arg Gln Glu Lys Ser Ser Gln Gln Asn Ser Gly Ala Arg 260 265270 Leu Pro Gly Leu Asn Lys Asn Gln Ser Gln Ser Gln Asp Ala Leu Val 275280 285 Leu Glu Asp Val Glu Lys Lys Lys Lys Lys Ser Gly Thr Lys Lys Asp290 295 300 Val Pro Lys Ser Trp Leu Met Lys Ala Leu Phe Lys Thr Phe TyrMet 305 310 315 320 Val Leu Leu Lys Ser Phe Leu Leu Lys Leu Val Asn AspIle Phe Thr 325 330 335 Phe Val Ser Pro Gln Leu Leu Lys Leu Leu Ile SerPhe Ala Ser Asp 340 345 350 Arg Asp Thr Tyr Leu Trp Ile Gly Tyr Leu CysAla Ile Leu Leu Phe 355 360 365 Thr Ala Ala Leu Ile Gln Ser Phe Cys LeuGln Cys Tyr Phe Gln Leu 370 375 380 Cys Phe Lys Leu Gly Val Lys Val ArgThr Ala Ile Met Ala Ser Val 385 390 395 400 Tyr Lys Lys Ala Leu Thr LeuSer Asn Leu Ala Arg Lys Glu Tyr Thr 405 410 415 Val Gly Glu Thr Val AsnLeu Met Ser Val Asp Ala Gln Lys Leu Met 420 425 430 Asp Val Thr Asn PheMet His Met Leu Trp Ser Ser Val Leu Gln Ile 435 440 445 Val Leu Ser IlePhe Phe Leu Trp Arg Glu Leu Gly Pro Ser Val Leu 450 455 460 Ala Gly ValGly Val Met Val Leu Val Ile Pro Ile Asn Ala Ile Leu 465 470 475 480 SerThr Lys Ser Lys Thr Ile Gln Val Lys Asn Met Lys Asn Lys Asp 485 490 495Lys Arg Leu Lys Ile Met Asn Glu Ile Leu Ser Gly Ile Lys Ile Leu 500 505510 Lys Tyr Phe Ala Trp Glu Pro Ser Phe Arg Asp Gln Val Gln Asn Leu 515520 525 Arg Lys Lys Glu Leu Lys Asn Leu Leu Ala Phe Ser Gln Leu Gln Cys530 535 540 Val Val Ile Phe Val Phe Gln Leu Thr Pro Val Leu Val Ser ValVal 545 550 555 560 Thr Phe Ser Val Tyr Val Leu Val Asp Ser Asn Asn IleLeu Asp Ala 565 570 575 Gln Lys Ala Phe Thr Ser Ile Thr Leu Phe Asn IleLeu Arg Phe Pro 580 585 590 Leu Ser Met Leu Pro Met Met Ile Ser Ser MetLeu Gln Ala Ser Val 595 600 605 Ser Thr Glu Arg Leu Glu Lys Tyr Leu GlyGly Asp Asp Leu Asp Thr 610 615 620 Ser Ala Ile Arg His Asp Cys Asn PheAsp Lys Ala Met Gln Phe Ser 625 630 635 640 Glu Ala Ser Phe Thr Trp GluHis Asp Ser Glu Ala Thr Val Arg Asp 645 650 655 Val Asn Leu Asp Ile MetAla Gly Gln Leu Val Ala Val Ile Gly Pro 660 665 670 Val Gly Ser Gly LysSer Ser Leu Ile Ser Ala Met Leu Gly Glu Met 675 680 685 Glu Asn Val HisGly His Ile Thr Ile Lys Gly Thr Thr Ala Tyr Val 690 695 700 Pro Gln GlnSer Trp Ile Gln Asn Gly Thr Ile Lys Asp Asn Ile Leu 705 710 715 720 PheGly Thr Glu Phe Asn Glu Lys Arg Tyr Gln Gln Val Leu Glu Ala 725 730 735Cys Ala Leu Leu Pro Asp Leu Glu Met Leu Pro Gly Gly Asp Leu Ala 740 745750 Glu Ile Gly Glu Lys Gly Ile Asn Leu Ser Gly Gly Gln Lys Gln Arg 755760 765 Ile Ser Leu Ala Arg Ala Thr Tyr Gln Asn Leu Asp Ile Tyr Leu Leu770 775 780 Asp Asp Pro Leu Ser Ala Val Asp Ala His Val Gly Lys His IlePhe 785 790 795 800 Asn Lys Val Leu Gly Pro Asn Gly Leu Leu Lys Gly LysThr Arg Leu 805 810 815 Leu Val Thr His Ser Met His Phe Leu Pro Gln ValAsp Glu Ile Val 820 825 830 Val Leu Gly Asn Gly Thr Ile Val Glu Lys GlySer Tyr Ser Ala Leu 835 840 845 Leu Ala Lys Lys Gly Glu Phe Ala Lys AsnLeu Lys Thr Phe Leu Arg 850 855 860 His Thr Gly Pro Glu Glu Glu Ala ThrVal His Asp Gly Ser Glu Glu 865 870 875 880 Glu Asp Asp Asp Tyr Gly LeuIle Ser Ser Val Glu Glu Ile Pro Glu 885 890 895 Asp Ala Ala Ser Ile ThrMet Arg Arg Glu Asn Ser Phe Arg Arg Thr 900 905 910 Leu Ser Arg Ser SerArg Ser Asn Gly Arg His Leu Lys Ser Leu Arg 915 920 925 Asn Ser Leu LysThr Arg Asn Val Asn Ser Leu Lys Glu Asp Glu Glu 930 935 940 Leu Val LysGly Gln Lys Leu Ile Lys Lys Glu Phe Ile Glu Thr Gly 945 950 955 960 LysVal Lys Phe Ser Ile Tyr Leu Glu Tyr Leu Gln Ala Ile Gly Leu 965 970 975Phe Ser Ile Phe Phe Ile Ile Leu Ala Phe Val Met Asn Ser Val Ala 980 985990 Phe Ile Gly Ser Asn Leu Trp Leu Ser Ala Trp Thr Ser Asp Ser Lys 9951000 1005 Ile Phe Asn Ser Thr Asp Tyr Pro Ala Ser Gln Arg Asp Met Arg1010 1015 1020 Val Gly Val Tyr Gly Ala Leu Gly Leu Ala Gln Gly Ile PheVal 1025 1030 1035 Phe Ile Ala His Phe Trp Ser Ala Phe Gly Phe Val HisAla Ser 1040 1045 1050 Asn Ile Leu His Lys Gln Leu Leu Asn Asn Ile LeuArg Ala Pro 1055 1060 1065 Met Arg Phe Phe Asp Thr Thr Pro Thr Gly ArgIle Val Asn Arg 1070 1075 1080 Phe Ala Gly Asp Ile Ser Thr Val Asp AspThr Leu Pro Gln Ser 1085 1090 1095 Leu Arg Ser Trp Ile Thr Cys Phe LeuGly Ile Ile Ser Thr Leu 1100 1105 1110 Val Met Ile Cys Met Ala Thr ProVal Phe Thr Ile Ile Val Ile 1115 1120 1125 Pro Leu Gly Ile Ile Tyr ValSer Val Gln Met Phe Tyr Val Ser 1130 1135 1140 Thr Ser Arg Gln Leu ArgArg Leu Asp Ser Val Thr Arg Ser Pro 1145 1150 1155 Ile Tyr Ser His PheSer Glu Thr Val Ser Gly Leu Pro Val Ile 1160 1165 1170 Arg Ala Phe GluHis Gln Gln Arg Phe Leu Lys His Asn Glu Glu 1175 1180 1185 Arg Ile AspThr Asn Gln Lys Cys Val Phe Ser Trp Ile Thr Ser 1190 1195 1200 Asn ArgTrp Leu Ala Ile Arg Leu Glu Leu Val Gly Asn Leu Thr 1205 1210 1215 ValPhe Phe Ser Ala Leu Met Met Val Ile Tyr Arg Asp Thr Leu 1220 1225 1230Ser Gly Asp Thr Val Gly Phe Val Leu Ser Asn Ala Leu Asn Ile 1235 12401245 Thr Gln Thr Leu Asn Trp Leu Val Arg Met Thr Ser Glu Ile Glu 12501255 1260 Thr Asn Ile Val Ala Val Glu Arg Ile Thr Glu Tyr Thr Lys Val1265 1270 1275 Glu Asn Glu Ala Pro Trp Val Thr Asp Lys Arg Pro Pro ProAsp 1280 1285 1290 Trp Pro Ser Lys Gly Lys Ile Gln Phe Asn Asn Tyr GlnVal Arg 1295 1300 1305 Tyr Arg Pro Glu Leu Asp Leu Val Leu Arg Gly IleThr Cys Asp 1310 1315 1320 Ile Gly Ser Met Glu Lys Ile Gly Val Val GlyArg Thr Gly Ala 1325 1330 1335 Gly Lys Ser Ser Leu Thr Asn Cys Leu PheArg Ile Leu Glu Ala 1340 1345 1350 Ala Gly Gly Gln Ile Ile Ile Asp GlyVal Asp Ile Ala Ser Ile 1355 1360 1365 Gly Leu His Asp Leu Arg Glu LysLeu Thr Ile Ile Pro Gln Asp 1370 1375 1380 Pro Ile Leu Phe Ser Gly SerLeu Arg Met Asn Leu Asp Pro Phe 1385 1390 1395 Asn Asn Tyr Ser Asp GluGlu Ile Trp Lys Ala Leu Glu Leu Ala 1400 1405 1410 His Leu Lys Ser PheVal Ala Ser Leu Gln Leu Gly Leu Ser His 1415 1420 1425 Glu Val Thr GluAla Gly Gly Asn Leu Ser Ile Gly Gln Arg Gln 1430 1435 1440 Leu Leu CysLeu Gly Arg Ala Leu Leu Arg Lys Ser Lys Ile Leu 1445 1450 1455 Val LeuAsp Glu Ala Thr Ala Ala Val Asp Leu Glu Thr Asp Asn 1460 1465 1470 LeuIle Gln Thr Thr Ile Gln Asn Glu Phe Ala His Cys Thr Val 1475 1480 1485Ile Thr Ile Ala His Arg Leu His Thr Ile Met Asp Ser Asp Lys 1490 14951500 Val Met Val Leu Asp Asn Gly Lys Ile Ile Glu Tyr Gly Ser Pro 15051510 1515 Glu Glu Leu Leu Gln Ile Pro Gly Pro Phe Tyr Phe Met Ala Lys1520 1525 1530 Glu Ala Gly Ile Glu Asn Val Asn Ala Thr Lys Phe 1535 15401545 9 4640 DNA Homo sapiens CDS (1)..(4635) 9 atg ctg gag aag ttc tgcaac tct act ttt tgg aat tcc tca ttc ctg 48 Met Leu Glu Lys Phe Cys AsnSer Thr Phe Trp Asn Ser Ser Phe Leu 1 5 10 15 gac agt ccg gag gca gacctg cca ctt tgt ttt gag caa act gtt ctg 96 Asp Ser Pro Glu Ala Asp LeuPro Leu Cys Phe Glu Gln Thr Val Leu 20 25 30 gtg tgg att ccc ttg ggc ttccta tgg ctc ctg gcc ccc tgg cag ctt 144 Val Trp Ile Pro Leu Gly Phe LeuTrp Leu Leu Ala Pro Trp Gln Leu 35 40 45 ctc cac gtg tat aaa tcc agg accaag aga tcc tct acc acc aaa ctc 192 Leu His Val Tyr Lys Ser Arg Thr LysArg Ser Ser Thr Thr Lys Leu 50 55 60 tat ctt gct aag cag gta ttc gtt ggtttt ctt ctt att cta gca gcc 240 Tyr Leu Ala Lys Gln Val Phe Val Gly PheLeu Leu Ile Leu Ala Ala 65 70 75 80 ata gag ctg gcc ctt gta ctc aca gaagac tct gga caa gcc aca gtc 288 Ile Glu Leu Ala Leu Val Leu Thr Glu AspSer Gly Gln Ala Thr Val 85 90 95 cct gct gtt cga tat acc aat cca agc ctctac cta ggc aca tgg ctc 336 Pro Ala Val Arg Tyr Thr Asn Pro Ser Leu TyrLeu Gly Thr Trp Leu 100 105 110 ctg gtt ttg ctg atc caa tac agc aga caatgg tgt gta cag aaa aac 384 Leu Val Leu Leu Ile Gln Tyr Ser Arg Gln TrpCys Val Gln Lys Asn 115 120 125 tcc tgg ttc ctg tcc cta ttc tgg att ctctcg ata ctc tgt ggc act 432 Ser Trp Phe Leu Ser Leu Phe Trp Ile Leu SerIle Leu Cys Gly Thr 130 135 140 ttc caa ttt cag act ctg atc cgg aca ctctta cag ggt gac aat tct 480 Phe Gln Phe Gln Thr Leu Ile Arg Thr Leu LeuGln Gly Asp Asn Ser 145 150 155 160 aat cta gcc tac tcc tgc ctg ttc ttcatc tcc tac gga ttc cag atc 528 Asn Leu Ala Tyr Ser Cys Leu Phe Phe IleSer Tyr Gly Phe Gln Ile 165 170 175 ctg atc ctg atc ttt tca gca ttt tcagaa aat aat gag tca tca aat 576 Leu Ile Leu Ile Phe Ser Ala Phe Ser GluAsn Asn Glu Ser Ser Asn 180 185 190 aat cca tca tcc ata gct tca ttc ctgagt agc att acc tac agc tgg 624 Asn Pro Ser Ser Ile Ala Ser Phe Leu SerSer Ile Thr Tyr Ser Trp 195 200 205 tat gac agc atc att ctg aaa ggc tacaag cgt cct ctg aca ctc gag 672 Tyr Asp Ser Ile Ile Leu Lys Gly Tyr LysArg Pro Leu Thr Leu Glu 210 215 220 gat gtc tgg gaa gtt gat gaa gag atgaaa acc aag aca tta gtg agc 720 Asp Val Trp Glu Val Asp Glu Glu Met LysThr Lys Thr Leu Val Ser 225 230 235 240 aag ttt gaa acg cac atg aag agagag ctg cag aaa gcc agg cgg gca 768 Lys Phe Glu Thr His Met Lys Arg GluLeu Gln Lys Ala Arg Arg Ala 245 250 255 ctc cag aga cgg cag gag aag agctcc cag cag aac tct gga gcc agg 816 Leu Gln Arg Arg Gln Glu Lys Ser SerGln Gln Asn Ser Gly Ala Arg 260 265 270 ctg cct ggc ttg aac aag aat cagagt caa agc caa gat gcc ctt gtc 864 Leu Pro Gly Leu Asn Lys Asn Gln SerGln Ser Gln Asp Ala Leu Val 275 280 285 ctg gaa gat gtt gaa aag aaa aaaaag aag tct ggg acc aaa aaa gat 912 Leu Glu Asp Val Glu Lys Lys Lys LysLys Ser Gly Thr Lys Lys Asp 290 295 300 gtt cca aaa tcc tgg ttg atg aaggct ctg ttc aaa act ttc tac atg 960 Val Pro Lys Ser Trp Leu Met Lys AlaLeu Phe Lys Thr Phe Tyr Met 305 310 315 320 gtg ctc ctg aaa tca ttc ctactg aag cta gtg aat gac atc ttc acg 1008 Val Leu Leu Lys Ser Phe Leu LeuLys Leu Val Asn Asp Ile Phe Thr 325 330 335 ttt gtg agt cct cag ctg ctgaaa ttg ctg atc tcc ttt gca agt gac 1056 Phe Val Ser Pro Gln Leu Leu LysLeu Leu Ile Ser Phe Ala Ser Asp 340 345 350 cgt gac aca tat ttg tgg attgga tat ctc tgt gca atc ctc tta ttc 1104 Arg Asp Thr Tyr Leu Trp Ile GlyTyr Leu Cys Ala Ile Leu Leu Phe 355 360 365 act gcg gct ctc att cag tctttc tgc ctt cag tgt tat ttc caa ctg 1152 Thr Ala Ala Leu Ile Gln Ser PheCys Leu Gln Cys Tyr Phe Gln Leu 370 375 380 tgc ttc aag ctg ggt gta aaagta cgg aca gct atc atg gct tct gta 1200 Cys Phe Lys Leu Gly Val Lys ValArg Thr Ala Ile Met Ala Ser Val 385 390 395 400 tat aag aag gca ttg acccta tcc aac ttg gcc agg aag gag tac acc 1248 Tyr Lys Lys Ala Leu Thr LeuSer Asn Leu Ala Arg Lys Glu Tyr Thr 405 410 415 gtt gga gaa aca gtg aacctg atg tct gtg gat gcc cag aag ctc atg 1296 Val Gly Glu Thr Val Asn LeuMet Ser Val Asp Ala Gln Lys Leu Met 420 425 430 gat gtg acc aac ttc atgcac atg ctg tgg tca agt gtt cta cag att 1344 Asp Val Thr Asn Phe Met HisMet Leu Trp Ser Ser Val Leu Gln Ile 435 440 445 gtc tta tct atc ttc ttccta tgg aga gag ttg gga ccc tca gtc tta 1392 Val Leu Ser Ile Phe Phe LeuTrp Arg Glu Leu Gly Pro Ser Val Leu 450 455 460 gca ggt gtt ggg gtg atggtg ctt gta atc cca att aat gcg ata ctg 1440 Ala Gly Val Gly Val Met ValLeu Val Ile Pro Ile Asn Ala Ile Leu 465 470 475 480 tcc acc aag agt aagacc att cag gtc aaa aat atg aag aat aaa gac 1488 Ser Thr Lys Ser Lys ThrIle Gln Val Lys Asn Met Lys Asn Lys Asp 485 490 495 aaa cgt tta aag atcatg aat gag att ctt agt gga atc aag atc ctg 1536 Lys Arg Leu Lys Ile MetAsn Glu Ile Leu Ser Gly Ile Lys Ile Leu 500 505 510 aaa tat ttt gcc tgggaa cct tca ttc aga gac caa gta caa aac ctc 1584 Lys Tyr Phe Ala Trp GluPro Ser Phe Arg Asp Gln Val Gln Asn Leu 515 520 525 cgg aag aaa gag ctcaag aac ctg ctg gcc ttt agt caa cta cag tgt 1632 Arg Lys Lys Glu Leu LysAsn Leu Leu Ala Phe Ser Gln Leu Gln Cys 530 535 540 gta gta ata ttc gtcttc cag tta act cca gtc ctg gta tct gtg gtc 1680 Val Val Ile Phe Val PheGln Leu Thr Pro Val Leu Val Ser Val Val 545 550 555 560 aca ttt tct gtttat gtc ctg gtg gat agc aac aat att ttg gat gca 1728 Thr Phe Ser Val TyrVal Leu Val Asp Ser Asn Asn Ile Leu Asp Ala 565 570 575 caa aag gcc ttcacc tcc att acc ctc ttc aat atc ctg cgc ttt ccc 1776 Gln Lys Ala Phe ThrSer Ile Thr Leu Phe Asn Ile Leu Arg Phe Pro 580 585 590 ctg agc atg cttccc atg atg atc tcc tcc atg ctc cag gcc agt gtt 1824 Leu Ser Met Leu ProMet Met Ile Ser Ser Met Leu Gln Ala Ser Val 595 600 605 tcc aca gag cggcta gag aag tac ttg gga ggg gat gac ttg gac aca 1872 Ser Thr Glu Arg LeuGlu Lys Tyr Leu Gly Gly Asp Asp Leu Asp Thr 610 615 620 tct gcc att cgacat gac tgc aat ttt gac aaa gcc atg cag ttt tct 1920 Ser Ala Ile Arg HisAsp Cys Asn Phe Asp Lys Ala Met Gln Phe Ser 625 630 635 640 gag gcc tccttt acc tgg gaa cat gat tcg gaa gcc aca gtc cga gat 1968 Glu Ala Ser PheThr Trp Glu His Asp Ser Glu Ala Thr Val Arg Asp 645 650 655 gtg aac ctggac att atg gca ggc caa ctt gtg gct gtg ata ggc cct 2016 Val Asn Leu AspIle Met Ala Gly Gln Leu Val Ala Val Ile Gly Pro 660 665 670 gtc ggc tctggg aaa tcc tcc ttg ata tca gcc atg ctg gga gaa atg 2064 Val Gly Ser GlyLys Ser Ser Leu Ile Ser Ala Met Leu Gly Glu Met 675 680 685 gaa aat gtccac ggg cac atc acc atc aag ggc acc act gcc tat gtc 2112 Glu Asn Val HisGly His Ile Thr Ile Lys Gly Thr Thr Ala Tyr Val 690 695 700 cca cag cagtcc tgg att cag aat ggc acc ata aag gac aac atc ctt 2160 Pro Gln Gln SerTrp Ile Gln Asn Gly Thr Ile Lys Asp Asn Ile Leu 705 710 715 720 ttt ggaaca gag ttt aat gaa aag agg tac cag caa gta ctg gag gcc 2208 Phe Gly ThrGlu Phe Asn Glu Lys Arg Tyr Gln Gln Val Leu Glu Ala 725 730 735 tgt gctctc ctc cca gac ttg gaa atg ctg cct gga gga gat ttg gct 2256 Cys Ala LeuLeu Pro Asp Leu Glu Met Leu Pro Gly Gly Asp Leu Ala 740 745 750 gag attgga gag aag ggt ata aat ctt agt ggg ggt cag aag cag cgg 2304 Glu Ile GlyGlu Lys Gly Ile Asn Leu Ser Gly Gly Gln Lys Gln Arg 755 760 765 atc agcctg gcc aga gct acc tac caa aat tta gac atc tat ctt cta 2352 Ile Ser LeuAla Arg Ala Thr Tyr Gln Asn Leu Asp Ile Tyr Leu Leu 770 775 780 gat gacccc ctg tct gca gtg gat gct cat gta gga aaa cat att ttt 2400 Asp Asp ProLeu Ser Ala Val Asp Ala His Val Gly Lys His Ile Phe 785 790 795 800 aataag gtc ttg ggc ccc aat ggc ctg ttg aaa ggc aag act cga ctc 2448 Asn LysVal Leu Gly Pro Asn Gly Leu Leu Lys Gly Lys Thr Arg Leu 805 810 815 ttggtt aca cat agc atg cac ttt ctt cct caa gtg gat gag att gta 2496 Leu ValThr His Ser Met His Phe Leu Pro Gln Val Asp Glu Ile Val 820 825 830 gttctg ggg aat gga aca att gta gag aaa gga tcc tac agt gct ctc 2544 Val LeuGly Asn Gly Thr Ile Val Glu Lys Gly Ser Tyr Ser Ala Leu 835 840 845 ctggcc aaa aaa gga gag ttt gct aag aat ctg aag aca ttt cta aga 2592 Leu AlaLys Lys Gly Glu Phe Ala Lys Asn Leu Lys Thr Phe Leu Arg 850 855 860 cataca ggc cct gaa gag gaa gcc aca gtc cat gat ggc agt gaa gaa 2640 His ThrGly Pro Glu Glu Glu Ala Thr Val His Asp Gly Ser Glu Glu 865 870 875 880gaa gac gat gac tat ggg ctg ata tcc agt gtg gaa gag atc ccc gaa 2688 GluAsp Asp Asp Tyr Gly Leu Ile Ser Ser Val Glu Glu Ile Pro Glu 885 890 895gat gca gcc tcc ata acc atg aga aga gag aac agc ttt cgt cga aca 2736 AspAla Ala Ser Ile Thr Met Arg Arg Glu Asn Ser Phe Arg Arg Thr 900 905 910ctt agc cgc agt tct agg tcc aat ggc agg cat ctg aag tcc ctg aga 2784 LeuSer Arg Ser Ser Arg Ser Asn Gly Arg His Leu Lys Ser Leu Arg 915 920 925aac tcc ttg aaa act cgg aat gtg aat agc ctg aag gaa gac gaa gaa 2832 AsnSer Leu Lys Thr Arg Asn Val Asn Ser Leu Lys Glu Asp Glu Glu 930 935 940cta gtg aaa gga caa aaa cta att aag aag gaa ttc ata gaa act gga 2880 LeuVal Lys Gly Gln Lys Leu Ile Lys Lys Glu Phe Ile Glu Thr Gly 945 950 955960 aag gtg aag ttc tcc atc tac ctg gag tac cta caa gca ata gga ttg 2928Lys Val Lys Phe Ser Ile Tyr Leu Glu Tyr Leu Gln Ala Ile Gly Leu 965 970975 ttt tcg ata ttc ttc atc atc ctt gcg ttt gtg atg aat tct gtg gct 2976Phe Ser Ile Phe Phe Ile Ile Leu Ala Phe Val Met Asn Ser Val Ala 980 985990 ttt att gga tcc aac ctc tgg ctc agt gct tgg acc agt gac tct aaa 3024Phe Ile Gly Ser Asn Leu Trp Leu Ser Ala Trp Thr Ser Asp Ser Lys 995 10001005 atc ttc aat agc acc gac tat cca gca tct cag agg gac atg aga 3069Ile Phe Asn Ser Thr Asp Tyr Pro Ala Ser Gln Arg Asp Met Arg 1010 10151020 gtt gga gtc tac gga gct ctg gga tta gcc caa ggt ata ttt gtg 3114Val Gly Val Tyr Gly Ala Leu Gly Leu Ala Gln Gly Ile Phe Val 1025 10301035 ttc ata gca cat ttc tgg agt gcc ttt ggt ttc gtc cat gca tca 3159Phe Ile Ala His Phe Trp Ser Ala Phe Gly Phe Val His Ala Ser 1040 10451050 aat atc ttg cac aag caa ctg ctg aac aat atc ctt cga gca cct 3204Asn Ile Leu His Lys Gln Leu Leu Asn Asn Ile Leu Arg Ala Pro 1055 10601065 atg aga ttt ttt gac aca aca ccc aca ggc cgg att gtg aac agg 3249Met Arg Phe Phe Asp Thr Thr Pro Thr Gly Arg Ile Val Asn Arg 1070 10751080 ttt gcc ggc gat att tcc aca gtg gat gac acc ctg cct cag tcc 3294Phe Ala Gly Asp Ile Ser Thr Val Asp Asp Thr Leu Pro Gln Ser 1085 10901095 ttg cgc agc tgg att aca tgc ttc ctg ggg ata atc agc acc ctt 3339Leu Arg Ser Trp Ile Thr Cys Phe Leu Gly Ile Ile Ser Thr Leu 1100 11051110 gtc atg atc tgc atg gcc act cct gtc ttc acc atc atc gtc att 3384Val Met Ile Cys Met Ala Thr Pro Val Phe Thr Ile Ile Val Ile 1115 11201125 cct ctt ggc att att tat gta tct gtt cag atg ttt tat gtg tct 3429Pro Leu Gly Ile Ile Tyr Val Ser Val Gln Met Phe Tyr Val Ser 1130 11351140 acc tcc cgc cag ctg agg cgt ctg gac tct gtc acc agg tcc cca 3474Thr Ser Arg Gln Leu Arg Arg Leu Asp Ser Val Thr Arg Ser Pro 1145 11501155 atc tac tct cac ttc agc gag acc gta tca ggt ttg cca gtt atc 3519Ile Tyr Ser His Phe Ser Glu Thr Val Ser Gly Leu Pro Val Ile 1160 11651170 cgt gcc ttt gag cac cag cag cga ttt ctg aaa cac aat gag gag 3564Arg Ala Phe Glu His Gln Gln Arg Phe Leu Lys His Asn Glu Glu 1175 11801185 agg att gac acc aac cag aaa tgt gtc ttt tcc tgg atc acc tcc 3609Arg Ile Asp Thr Asn Gln Lys Cys Val Phe Ser Trp Ile Thr Ser 1190 11951200 aac agg tgg ctt gca att cgc ctg gag ctg gtt ggg aac ctg act 3654Asn Arg Trp Leu Ala Ile Arg Leu Glu Leu Val Gly Asn Leu Thr 1205 12101215 gtc ttc ttt tca gcc ttg atg atg gtt att tat aga gat acc cta 3699Val Phe Phe Ser Ala Leu Met Met Val Ile Tyr Arg Asp Thr Leu 1220 12251230 agt ggg gac act gtt ggc ttt gtt ctg tcc aat gca ctc aat atc 3744Ser Gly Asp Thr Val Gly Phe Val Leu Ser Asn Ala Leu Asn Ile 1235 12401245 aca caa acc ctg aac tgg ctg gtg agg atg aca tca gaa ata gag 3789Thr Gln Thr Leu Asn Trp Leu Val Arg Met Thr Ser Glu Ile Glu 1250 12551260 acc aac att gtg gct gtt gag cga ata act gag tac aca aaa gtg 3834Thr Asn Ile Val Ala Val Glu Arg Ile Thr Glu Tyr Thr Lys Val 1265 12701275 gaa aat gag gca ccc tgg gtg act gat aag agg cct ccg cca gat 3879Glu Asn Glu Ala Pro Trp Val Thr Asp Lys Arg Pro Pro Pro Asp 1280 12851290 tgg ccc agc aaa ggc aag atc cag ttt aac aac tac caa gtg cgg 3924Trp Pro Ser Lys Gly Lys Ile Gln Phe Asn Asn Tyr Gln Val Arg 1295 13001305 tac cga cct gag ctg gat ctg gtc ctc aga ggg atc act tgt gac 3969Tyr Arg Pro Glu Leu Asp Leu Val Leu Arg Gly Ile Thr Cys Asp 1310 13151320 atc ggt agc atg gag aag att ggt gtg gtg ggc agg aca gga gct 4014Ile Gly Ser Met Glu Lys Ile Gly Val Val Gly Arg Thr Gly Ala 1325 13301335 gga aag tca tcc ctc aca aac tgc ctc ttc aga atc tta gag gct 4059Gly Lys Ser Ser Leu Thr Asn Cys Leu Phe Arg Ile Leu Glu Ala 1340 13451350 gcc ggt ggt cag att atc att gat gga gta gat att gct tcc att 4104Ala Gly Gly Gln Ile Ile Ile Asp Gly Val Asp Ile Ala Ser Ile 1355 13601365 ggg ctc cac gac ctc cga gag aag ctg acc atc atc ccc cag gac 4149Gly Leu His Asp Leu Arg Glu Lys Leu Thr Ile Ile Pro Gln Asp 1370 13751380 ccc atc ctg ttc tct gga agc ctg agg atg aat ctc gac cct ttc 4194Pro Ile Leu Phe Ser Gly Ser Leu Arg Met Asn Leu Asp Pro Phe 1385 13901395 aac aac tac tca gat gag gag att tgg aag gcc ttg gag ctg gct 4239Asn Asn Tyr Ser Asp Glu Glu Ile Trp Lys Ala Leu Glu Leu Ala 1400 14051410 cac ctc aag tct ttt gtg gcc agc ctg caa ctt ggg tta tcc cac 4284His Leu Lys Ser Phe Val Ala Ser Leu Gln Leu Gly Leu Ser His 1415 14201425 gaa gtt aca gag gct ggt ggc aac ctg agc ata ggc cag agg cag 4329Glu Val Thr Glu Ala Gly Gly Asn Leu Ser Ile Gly Gln Arg Gln 1430 14351440 ctg ctg tgc ctg ggc agg gct ctg ctt cgg aaa tcc aag atc ctg 4374Leu Leu Cys Leu Gly Arg Ala Leu Leu Arg Lys Ser Lys Ile Leu 1445 14501455 gtc ctg gat gag gcc act gct gcg gtg gat cta gag aca gac aac 4419Val Leu Asp Glu Ala Thr Ala Ala Val Asp Leu Glu Thr Asp Asn 1460 14651470 ctc att cag acg acc atc caa aac gag ttc gcc cac tgc aca gtg 4464Leu Ile Gln Thr Thr Ile Gln Asn Glu Phe Ala His Cys Thr Val 1475 14801485 atc acc atc gcc cac agg ctg cat acc atc atg gac agt gac aag 4509Ile Thr Ile Ala His Arg Leu His Thr Ile Met Asp Ser Asp Lys 1490 14951500 gta atg gtc cta gac aac ggg aag att ata gag tac ggc agc cct 4554Val Met Val Leu Asp Asn Gly Lys Ile Ile Glu Tyr Gly Ser Pro 1505 15101515 gaa gaa ctg cta caa atc cct gga ccc ttt tac ttt atg gct aag 4599Glu Glu Leu Leu Gln Ile Pro Gly Pro Phe Tyr Phe Met Ala Lys 1520 15251530 gaa gct ggc att gag aat gtg aac agc gca aaa ttc gcacc 4640 Glu AlaGly Ile Glu Asn Val Asn Ser Ala Lys Phe 1535 1540 1545 10 1545 PRT Homosapiens 10 Met Leu Glu Lys Phe Cys Asn Ser Thr Phe Trp Asn Ser Ser PheLeu 1 5 10 15 Asp Ser Pro Glu Ala Asp Leu Pro Leu Cys Phe Glu Gln ThrVal Leu 20 25 30 Val Trp Ile Pro Leu Gly Phe Leu Trp Leu Leu Ala Pro TrpGln Leu 35 40 45 Leu His Val Tyr Lys Ser Arg Thr Lys Arg Ser Ser Thr ThrLys Leu 50 55 60 Tyr Leu Ala Lys Gln Val Phe Val Gly Phe Leu Leu Ile LeuAla Ala 65 70 75 80 Ile Glu Leu Ala Leu Val Leu Thr Glu Asp Ser Gly GlnAla Thr Val 85 90 95 Pro Ala Val Arg Tyr Thr Asn Pro Ser Leu Tyr Leu GlyThr Trp Leu 100 105 110 Leu Val Leu Leu Ile Gln Tyr Ser Arg Gln Trp CysVal Gln Lys Asn 115 120 125 Ser Trp Phe Leu Ser Leu Phe Trp Ile Leu SerIle Leu Cys Gly Thr 130 135 140 Phe Gln Phe Gln Thr Leu Ile Arg Thr LeuLeu Gln Gly Asp Asn Ser 145 150 155 160 Asn Leu Ala Tyr Ser Cys Leu PhePhe Ile Ser Tyr Gly Phe Gln Ile 165 170 175 Leu Ile Leu Ile Phe Ser AlaPhe Ser Glu Asn Asn Glu Ser Ser Asn 180 185 190 Asn Pro Ser Ser Ile AlaSer Phe Leu Ser Ser Ile Thr Tyr Ser Trp 195 200 205 Tyr Asp Ser Ile IleLeu Lys Gly Tyr Lys Arg Pro Leu Thr Leu Glu 210 215 220 Asp Val Trp GluVal Asp Glu Glu Met Lys Thr Lys Thr Leu Val Ser 225 230 235 240 Lys PheGlu Thr His Met Lys Arg Glu Leu Gln Lys Ala Arg Arg Ala 245 250 255 LeuGln Arg Arg Gln Glu Lys Ser Ser Gln Gln Asn Ser Gly Ala Arg 260 265 270Leu Pro Gly Leu Asn Lys Asn Gln Ser Gln Ser Gln Asp Ala Leu Val 275 280285 Leu Glu Asp Val Glu Lys Lys Lys Lys Lys Ser Gly Thr Lys Lys Asp 290295 300 Val Pro Lys Ser Trp Leu Met Lys Ala Leu Phe Lys Thr Phe Tyr Met305 310 315 320 Val Leu Leu Lys Ser Phe Leu Leu Lys Leu Val Asn Asp IlePhe Thr 325 330 335 Phe Val Ser Pro Gln Leu Leu Lys Leu Leu Ile Ser PheAla Ser Asp 340 345 350 Arg Asp Thr Tyr Leu Trp Ile Gly Tyr Leu Cys AlaIle Leu Leu Phe 355 360 365 Thr Ala Ala Leu Ile Gln Ser Phe Cys Leu GlnCys Tyr Phe Gln Leu 370 375 380 Cys Phe Lys Leu Gly Val Lys Val Arg ThrAla Ile Met Ala Ser Val 385 390 395 400 Tyr Lys Lys Ala Leu Thr Leu SerAsn Leu Ala Arg Lys Glu Tyr Thr 405 410 415 Val Gly Glu Thr Val Asn LeuMet Ser Val Asp Ala Gln Lys Leu Met 420 425 430 Asp Val Thr Asn Phe MetHis Met Leu Trp Ser Ser Val Leu Gln Ile 435 440 445 Val Leu Ser Ile PhePhe Leu Trp Arg Glu Leu Gly Pro Ser Val Leu 450 455 460 Ala Gly Val GlyVal Met Val Leu Val Ile Pro Ile Asn Ala Ile Leu 465 470 475 480 Ser ThrLys Ser Lys Thr Ile Gln Val Lys Asn Met Lys Asn Lys Asp 485 490 495 LysArg Leu Lys Ile Met Asn Glu Ile Leu Ser Gly Ile Lys Ile Leu 500 505 510Lys Tyr Phe Ala Trp Glu Pro Ser Phe Arg Asp Gln Val Gln Asn Leu 515 520525 Arg Lys Lys Glu Leu Lys Asn Leu Leu Ala Phe Ser Gln Leu Gln Cys 530535 540 Val Val Ile Phe Val Phe Gln Leu Thr Pro Val Leu Val Ser Val Val545 550 555 560 Thr Phe Ser Val Tyr Val Leu Val Asp Ser Asn Asn Ile LeuAsp Ala 565 570 575 Gln Lys Ala Phe Thr Ser Ile Thr Leu Phe Asn Ile LeuArg Phe Pro 580 585 590 Leu Ser Met Leu Pro Met Met Ile Ser Ser Met LeuGln Ala Ser Val 595 600 605 Ser Thr Glu Arg Leu Glu Lys Tyr Leu Gly GlyAsp Asp Leu Asp Thr 610 615 620 Ser Ala Ile Arg His Asp Cys Asn Phe AspLys Ala Met Gln Phe Ser 625 630 635 640 Glu Ala Ser Phe Thr Trp Glu HisAsp Ser Glu Ala Thr Val Arg Asp 645 650 655 Val Asn Leu Asp Ile Met AlaGly Gln Leu Val Ala Val Ile Gly Pro 660 665 670 Val Gly Ser Gly Lys SerSer Leu Ile Ser Ala Met Leu Gly Glu Met 675 680 685 Glu Asn Val His GlyHis Ile Thr Ile Lys Gly Thr Thr Ala Tyr Val 690 695 700 Pro Gln Gln SerTrp Ile Gln Asn Gly Thr Ile Lys Asp Asn Ile Leu 705 710 715 720 Phe GlyThr Glu Phe Asn Glu Lys Arg Tyr Gln Gln Val Leu Glu Ala 725 730 735 CysAla Leu Leu Pro Asp Leu Glu Met Leu Pro Gly Gly Asp Leu Ala 740 745 750Glu Ile Gly Glu Lys Gly Ile Asn Leu Ser Gly Gly Gln Lys Gln Arg 755 760765 Ile Ser Leu Ala Arg Ala Thr Tyr Gln Asn Leu Asp Ile Tyr Leu Leu 770775 780 Asp Asp Pro Leu Ser Ala Val Asp Ala His Val Gly Lys His Ile Phe785 790 795 800 Asn Lys Val Leu Gly Pro Asn Gly Leu Leu Lys Gly Lys ThrArg Leu 805 810 815 Leu Val Thr His Ser Met His Phe Leu Pro Gln Val AspGlu Ile Val 820 825 830 Val Leu Gly Asn Gly Thr Ile Val Glu Lys Gly SerTyr Ser Ala Leu 835 840 845 Leu Ala Lys Lys Gly Glu Phe Ala Lys Asn LeuLys Thr Phe Leu Arg 850 855 860 His Thr Gly Pro Glu Glu Glu Ala Thr ValHis Asp Gly Ser Glu Glu 865 870 875 880 Glu Asp Asp Asp Tyr Gly Leu IleSer Ser Val Glu Glu Ile Pro Glu 885 890 895 Asp Ala Ala Ser Ile Thr MetArg Arg Glu Asn Ser Phe Arg Arg Thr 900 905 910 Leu Ser Arg Ser Ser ArgSer Asn Gly Arg His Leu Lys Ser Leu Arg 915 920 925 Asn Ser Leu Lys ThrArg Asn Val Asn Ser Leu Lys Glu Asp Glu Glu 930 935 940 Leu Val Lys GlyGln Lys Leu Ile Lys Lys Glu Phe Ile Glu Thr Gly 945 950 955 960 Lys ValLys Phe Ser Ile Tyr Leu Glu Tyr Leu Gln Ala Ile Gly Leu 965 970 975 PheSer Ile Phe Phe Ile Ile Leu Ala Phe Val Met Asn Ser Val Ala 980 985 990Phe Ile Gly Ser Asn Leu Trp Leu Ser Ala Trp Thr Ser Asp Ser Lys 995 10001005 Ile Phe Asn Ser Thr Asp Tyr Pro Ala Ser Gln Arg Asp Met Arg 10101015 1020 Val Gly Val Tyr Gly Ala Leu Gly Leu Ala Gln Gly Ile Phe Val1025 1030 1035 Phe Ile Ala His Phe Trp Ser Ala Phe Gly Phe Val His AlaSer 1040 1045 1050 Asn Ile Leu His Lys Gln Leu Leu Asn Asn Ile Leu ArgAla Pro 1055 1060 1065 Met Arg Phe Phe Asp Thr Thr Pro Thr Gly Arg IleVal Asn Arg 1070 1075 1080 Phe Ala Gly Asp Ile Ser Thr Val Asp Asp ThrLeu Pro Gln Ser 1085 1090 1095 Leu Arg Ser Trp Ile Thr Cys Phe Leu GlyIle Ile Ser Thr Leu 1100 1105 1110 Val Met Ile Cys Met Ala Thr Pro ValPhe Thr Ile Ile Val Ile 1115 1120 1125 Pro Leu Gly Ile Ile Tyr Val SerVal Gln Met Phe Tyr Val Ser 1130 1135 1140 Thr Ser Arg Gln Leu Arg ArgLeu Asp Ser Val Thr Arg Ser Pro 1145 1150 1155 Ile Tyr Ser His Phe SerGlu Thr Val Ser Gly Leu Pro Val Ile 1160 1165 1170 Arg Ala Phe Glu HisGln Gln Arg Phe Leu Lys His Asn Glu Glu 1175 1180 1185 Arg Ile Asp ThrAsn Gln Lys Cys Val Phe Ser Trp Ile Thr Ser 1190 1195 1200 Asn Arg TrpLeu Ala Ile Arg Leu Glu Leu Val Gly Asn Leu Thr 1205 1210 1215 Val PhePhe Ser Ala Leu Met Met Val Ile Tyr Arg Asp Thr Leu 1220 1225 1230 SerGly Asp Thr Val Gly Phe Val Leu Ser Asn Ala Leu Asn Ile 1235 1240 1245Thr Gln Thr Leu Asn Trp Leu Val Arg Met Thr Ser Glu Ile Glu 1250 12551260 Thr Asn Ile Val Ala Val Glu Arg Ile Thr Glu Tyr Thr Lys Val 12651270 1275 Glu Asn Glu Ala Pro Trp Val Thr Asp Lys Arg Pro Pro Pro Asp1280 1285 1290 Trp Pro Ser Lys Gly Lys Ile Gln Phe Asn Asn Tyr Gln ValArg 1295 1300 1305 Tyr Arg Pro Glu Leu Asp Leu Val Leu Arg Gly Ile ThrCys Asp 1310 1315 1320 Ile Gly Ser Met Glu Lys Ile Gly Val Val Gly ArgThr Gly Ala 1325 1330 1335 Gly Lys Ser Ser Leu Thr Asn Cys Leu Phe ArgIle Leu Glu Ala 1340 1345 1350 Ala Gly Gly Gln Ile Ile Ile Asp Gly ValAsp Ile Ala Ser Ile 1355 1360 1365 Gly Leu His Asp Leu Arg Glu Lys LeuThr Ile Ile Pro Gln Asp 1370 1375 1380 Pro Ile Leu Phe Ser Gly Ser LeuArg Met Asn Leu Asp Pro Phe 1385 1390 1395 Asn Asn Tyr Ser Asp Glu GluIle Trp Lys Ala Leu Glu Leu Ala 1400 1405 1410 His Leu Lys Ser Phe ValAla Ser Leu Gln Leu Gly Leu Ser His 1415 1420 1425 Glu Val Thr Glu AlaGly Gly Asn Leu Ser Ile Gly Gln Arg Gln 1430 1435 1440 Leu Leu Cys LeuGly Arg Ala Leu Leu Arg Lys Ser Lys Ile Leu 1445 1450 1455 Val Leu AspGlu Ala Thr Ala Ala Val Asp Leu Glu Thr Asp Asn 1460 1465 1470 Leu IleGln Thr Thr Ile Gln Asn Glu Phe Ala His Cys Thr Val 1475 1480 1485 IleThr Ile Ala His Arg Leu His Thr Ile Met Asp Ser Asp Lys 1490 1495 1500Val Met Val Leu Asp Asn Gly Lys Ile Ile Glu Tyr Gly Ser Pro 1505 15101515 Glu Glu Leu Leu Gln Ile Pro Gly Pro Phe Tyr Phe Met Ala Lys 15201525 1530 Glu Ala Gly Ile Glu Asn Val Asn Ser Ala Lys Phe 1535 1540 154511 4641 DNA Homo sapiens CDS (1)..(4635) 11 atg ctg gag aag ttc tgc aactct act ttt tgg aat tcc tca ttc ctg 48 Met Leu Glu Lys Phe Cys Asn SerThr Phe Trp Asn Ser Ser Phe Leu 1 5 10 15 gac agt ccg gag gca gac ctgcca ctt tgt ttt gag caa act gtt ctg 96 Asp Ser Pro Glu Ala Asp Leu ProLeu Cys Phe Glu Gln Thr Val Leu 20 25 30 gtg tgg att ccc ttg ggc ttc ctatgg ctc ctg gcc ccc tgg cag ctt 144 Val Trp Ile Pro Leu Gly Phe Leu TrpLeu Leu Ala Pro Trp Gln Leu 35 40 45 ctc cac gtg tat aaa tcc agg acc aagaga tcc tct acc acc aaa ctc 192 Leu His Val Tyr Lys Ser Arg Thr Lys ArgSer Ser Thr Thr Lys Leu 50 55 60 tat ctt gct aag cag gta ttc gtt ggt tttctt ctt att cta gca gcc 240 Tyr Leu Ala Lys Gln Val Phe Val Gly Phe LeuLeu Ile Leu Ala Ala 65 70 75 80 ata gag ctg gcc ctt gta ctc aca gaa gactct gga caa gcc aca gtc 288 Ile Glu Leu Ala Leu Val Leu Thr Glu Asp SerGly Gln Ala Thr Val 85 90 95 cct gct gtt cga tat acc aat cca agc ctc taccta ggc aca tgg ctc 336 Pro Ala Val Arg Tyr Thr Asn Pro Ser Leu Tyr LeuGly Thr Trp Leu 100 105 110 ctg gtt ttg ctg atc caa tac agc aga caa tggtgt gta cag aaa aac 384 Leu Val Leu Leu Ile Gln Tyr Ser Arg Gln Trp CysVal Gln Lys Asn 115 120 125 tcc tgg ttc ctg tcc cta ttc tgg att ctc tcgata ctc tgt ggc act 432 Ser Trp Phe Leu Ser Leu Phe Trp Ile Leu Ser IleLeu Cys Gly Thr 130 135 140 ttc caa ttt cag act ctg atc cgg aca ctc ttacag ggt gac aat tct 480 Phe Gln Phe Gln Thr Leu Ile Arg Thr Leu Leu GlnGly Asp Asn Ser 145 150 155 160 aat cta gcc tac tcc tgc ctg ttc ttc atctcc tac gga ttc cag atc 528 Asn Leu Ala Tyr Ser Cys Leu Phe Phe Ile SerTyr Gly Phe Gln Ile 165 170 175 ctg atc ctg atc ttt tca gca ttt tca gaaaat aat gag tca tca aat 576 Leu Ile Leu Ile Phe Ser Ala Phe Ser Glu AsnAsn Glu Ser Ser Asn 180 185 190 aat cca tca tcc ata gct tca ttc ctg agtagc att acc tac agc tgg 624 Asn Pro Ser Ser Ile Ala Ser Phe Leu Ser SerIle Thr Tyr Ser Trp 195 200 205 tat gac agc atc att ctg aaa ggc tac aagcgt cct ctg aca ctc gag 672 Tyr Asp Ser Ile Ile Leu Lys Gly Tyr Lys ArgPro Leu Thr Leu Glu 210 215 220 gat gtc tgg gaa gtt gat gaa gag atg aaaacc aag aca tta gtg agc 720 Asp Val Trp Glu Val Asp Glu Glu Met Lys ThrLys Thr Leu Val Ser 225 230 235 240 aag ttt gaa acg cac atg aag aga gagctg cag aaa gcc agg cgg gca 768 Lys Phe Glu Thr His Met Lys Arg Glu LeuGln Lys Ala Arg Arg Ala 245 250 255 ctc cag aga cgg cag gag aag agc tcccag cag aac tct gga gcc agg 816 Leu Gln Arg Arg Gln Glu Lys Ser Ser GlnGln Asn Ser Gly Ala Arg 260 265 270 ctg cct ggc ttg aac aag aat cag agtcaa agc caa gat gcc ctt gtc 864 Leu Pro Gly Leu Asn Lys Asn Gln Ser GlnSer Gln Asp Ala Leu Val 275 280 285 ctg gaa gat gtt gaa aag aaa aaa aagaag tct ggg acc aaa aaa gat 912 Leu Glu Asp Val Glu Lys Lys Lys Lys LysSer Gly Thr Lys Lys Asp 290 295 300 gtt cca aaa tcc tgg ttg atg aag gctctg ttc aaa act ttc tac atg 960 Val Pro Lys Ser Trp Leu Met Lys Ala LeuPhe Lys Thr Phe Tyr Met 305 310 315 320 gtg ctc ctg aaa tca ttc cta ctgaag cta gtg aat gac atc ttc acg 1008 Val Leu Leu Lys Ser Phe Leu Leu LysLeu Val Asn Asp Ile Phe Thr 325 330 335 ttt gtg agt cct cag ctg ctg aaattg ctg atc tcc ttt gca agt gac 1056 Phe Val Ser Pro Gln Leu Leu Lys LeuLeu Ile Ser Phe Ala Ser Asp 340 345 350 cgt gac aca tat ttg tgg att ggatat ctc tgt gca atc ctc tta ttc 1104 Arg Asp Thr Tyr Leu Trp Ile Gly TyrLeu Cys Ala Ile Leu Leu Phe 355 360 365 act gcg gct ctc att cag tct ttctgc ctt cag tgt tat ttc caa ctg 1152 Thr Ala Ala Leu Ile Gln Ser Phe CysLeu Gln Cys Tyr Phe Gln Leu 370 375 380 tgc ttc aag ctg ggt gta aaa gtacgg aca gct atc atg gct tct gta 1200 Cys Phe Lys Leu Gly Val Lys Val ArgThr Ala Ile Met Ala Ser Val 385 390 395 400 tat aag aag gca ttg acc ctatcc aac ttg gcc agg aag gag tac acc 1248 Tyr Lys Lys Ala Leu Thr Leu SerAsn Leu Ala Arg Lys Glu Tyr Thr 405 410 415 gtt gga gaa aca gtg aac ctgatg tct gtg gat gcc cag aag ctc atg 1296 Val Gly Glu Thr Val Asn Leu MetSer Val Asp Ala Gln Lys Leu Met 420 425 430 gat gtg acc aac ttc atg cacatg ctg tgg tca agt gtt cta cag att 1344 Asp Val Thr Asn Phe Met His MetLeu Trp Ser Ser Val Leu Gln Ile 435 440 445 gtc tta tct atc ttc ttc ctatgg aga gag ttg gga ccc tca gtc tta 1392 Val Leu Ser Ile Phe Phe Leu TrpArg Glu Leu Gly Pro Ser Val Leu 450 455 460 gca ggt gtt ggg gtg atg gtgctt gta atc cca att aat gcg ata ctg 1440 Ala Gly Val Gly Val Met Val LeuVal Ile Pro Ile Asn Ala Ile Leu 465 470 475 480 tcc acc aag agt aag accatt cag gtc aaa aat atg aag aat aaa gac 1488 Ser Thr Lys Ser Lys Thr IleGln Val Lys Asn Met Lys Asn Lys Asp 485 490 495 aaa cgt tta aag atc atgaat gag att ctt agt gga atc aag atc ctg 1536 Lys Arg Leu Lys Ile Met AsnGlu Ile Leu Ser Gly Ile Lys Ile Leu 500 505 510 aaa tat ttt gcc tgg gaacct tca ttc aga gac caa gta caa aac ctc 1584 Lys Tyr Phe Ala Trp Glu ProSer Phe Arg Asp Gln Val Gln Asn Leu 515 520 525 cgg aag aaa gag ctc aagaac ctg ctg gcc ttt agt caa cta cag tgt 1632 Arg Lys Lys Glu Leu Lys AsnLeu Leu Ala Phe Ser Gln Leu Gln Cys 530 535 540 gta gta ata ttc gtc ttccag tta act cca gtc ctg gta tct gtg gtc 1680 Val Val Ile Phe Val Phe GlnLeu Thr Pro Val Leu Val Ser Val Val 545 550 555 560 aca ttt tct gtt tatgtc ctg gtg gat agc aac aat att ttg gat gca 1728 Thr Phe Ser Val Tyr ValLeu Val Asp Ser Asn Asn Ile Leu Asp Ala 565 570 575 caa aag gcc ttc acctcc att acc ctc ttc aat atc ctg cgc ttt ccc 1776 Gln Lys Ala Phe Thr SerIle Thr Leu Phe Asn Ile Leu Arg Phe Pro 580 585 590 ctg agc atg ctt cccatg atg atc tcc tcc atg ctc cag gcc agt gtt 1824 Leu Ser Met Leu Pro MetMet Ile Ser Ser Met Leu Gln Ala Ser Val 595 600 605 tcc aca gag cgg ctagag aag tac ttg gga ggg gat gac ttg gac aca 1872 Ser Thr Glu Arg Leu GluLys Tyr Leu Gly Gly Asp Asp Leu Asp Thr 610 615 620 tct gcc att cga catgac tgc aat ttt gac aaa gcc atg cag ttt tct 1920 Ser Ala Ile Arg His AspCys Asn Phe Asp Lys Ala Met Gln Phe Ser 625 630 635 640 gag gcc tcc tttacc tgg gaa cat gat tcg gaa gcc aca gtc cga gat 1968 Glu Ala Ser Phe ThrTrp Glu His Asp Ser Glu Ala Thr Val Arg Asp 645 650 655 gtg aac ctg gacatt atg gca ggc caa ctt gtg gct gtg ata ggc cct 2016 Val Asn Leu Asp IleMet Ala Gly Gln Leu Val Ala Val Ile Gly Pro 660 665 670 gtc ggc tct gggaaa tcc tcc ttg ata tca gcc atg ctg gga gaa atg 2064 Val Gly Ser Gly LysSer Ser Leu Ile Ser Ala Met Leu Gly Glu Met 675 680 685 gaa aat gtc cacggg cac atc acc atc aag ggc acc act gcc tat gtc 2112 Glu Asn Val His GlyHis Ile Thr Ile Lys Gly Thr Thr Ala Tyr Val 690 695 700 cca cag cag tcctgg att cag aat ggc acc ata aag gac aac atc ctt 2160 Pro Gln Gln Ser TrpIle Gln Asn Gly Thr Ile Lys Asp Asn Ile Leu 705 710 715 720 ttt gga acagag ttt aat gaa aag agg tac cag caa gta ctg gag gcc 2208 Phe Gly Thr GluPhe Asn Glu Lys Arg Tyr Gln Gln Val Leu Glu Ala 725 730 735 tgt gct ctcctc cca gac ttg gaa atg ctg cct gga gga gat ttg gct 2256 Cys Ala Leu LeuPro Asp Leu Glu Met Leu Pro Gly Gly Asp Leu Ala 740 745 750 gag att ggagag aag ggt ata aat ctt agt ggg ggt cag aag cag cgg 2304 Glu Ile Gly GluLys Gly Ile Asn Leu Ser Gly Gly Gln Lys Gln Arg 755 760 765 atc agc ctggcc aga gct acc tac caa aat tta gac atc tat ctt cta 2352 Ile Ser Leu AlaArg Ala Thr Tyr Gln Asn Leu Asp Ile Tyr Leu Leu 770 775 780 gat gac cccctg tct gca gtg gat gct cat gta gga aaa cat att ttt 2400 Asp Asp Pro LeuSer Ala Val Asp Ala His Val Gly Lys His Ile Phe 785 790 795 800 aat aaggtc ttg ggc ccc aat ggc ctg ttg aaa ggc aag act cga ctc 2448 Asn Lys ValLeu Gly Pro Asn Gly Leu Leu Lys Gly Lys Thr Arg Leu 805 810 815 ttg gttaca cat agc atg cac ttt ctt cct caa gtg gat gag att gta 2496 Leu Val ThrHis Ser Met His Phe Leu Pro Gln Val Asp Glu Ile Val 820 825 830 gtt ctgggg aat gga aca att gta gag aaa gga tcc tac agt gct ctc 2544 Val Leu GlyAsn Gly Thr Ile Val Glu Lys Gly Ser Tyr Ser Ala Leu 835 840 845 ctg gccaaa aaa gga gag ttt gct aag aat ctg aag aca ttt cta aga 2592 Leu Ala LysLys Gly Glu Phe Ala Lys Asn Leu Lys Thr Phe Leu Arg 850 855 860 cat acaggc cct gaa gag gaa gcc aca gtc cat gat ggc agt gaa gaa 2640 His Thr GlyPro Glu Glu Glu Ala Thr Val His Asp Gly Ser Glu Glu 865 870 875 880 gaagac gat gac tat ggg ctg ata tcc agt gtg gaa gag atc ccc gaa 2688 Glu AspAsp Asp Tyr Gly Leu Ile Ser Ser Val Glu Glu Ile Pro Glu 885 890 895 gatgca gcc tcc ata acc atg aga aga gag aac agc ttt cgt cga aca 2736 Asp AlaAla Ser Ile Thr Met Arg Arg Glu Asn Ser Phe Arg Arg Thr 900 905 910 cttagc cgc agt tct agg tcc aat ggc agg cat ctg aag tcc ctg aga 2784 Leu SerArg Ser Ser Arg Ser Asn Gly Arg His Leu Lys Ser Leu Arg 915 920 925 aactcc ttg aaa act cgg aat gtg aat agc ctg aag gaa gac gaa gaa 2832 Asn SerLeu Lys Thr Arg Asn Val Asn Ser Leu Lys Glu Asp Glu Glu 930 935 940 ctagtg aaa gga caa aaa cta att aag aag gaa ttc ata gaa act gga 2880 Leu ValLys Gly Gln Lys Leu Ile Lys Lys Glu Phe Ile Glu Thr Gly 945 950 955 960aag gtg aag ttc tcc atc tac ctg gag tac cta caa gca ata gga ttg 2928 LysVal Lys Phe Ser Ile Tyr Leu Glu Tyr Leu Gln Ala Ile Gly Leu 965 970 975ttt tcg ata ttc ttc atc atc ctt gcg ttt gtg atg aat tct gtg gct 2976 PheSer Ile Phe Phe Ile Ile Leu Ala Phe Val Met Asn Ser Val Ala 980 985 990ttt att gga tcc aac ctc tgg ctc agt gct tgg acc agt gac tct aaa 3024 PheIle Gly Ser Asn Leu Trp Leu Ser Ala Trp Thr Ser Asp Ser Lys 995 10001005 atc ttc aat agc acc gac tat cca gca tct cag agg gac atg aga 3069Ile Phe Asn Ser Thr Asp Tyr Pro Ala Ser Gln Arg Asp Met Arg 1010 10151020 gtt gga gtc tac gga gct ctg gga tta gcc caa ggt ata ttt gtg 3114Val Gly Val Tyr Gly Ala Leu Gly Leu Ala Gln Gly Ile Phe Val 1025 10301035 ttc ata gca cat ttc tgg agt gcc ttt ggt ttc gtc cat gca tca 3159Phe Ile Ala His Phe Trp Ser Ala Phe Gly Phe Val His Ala Ser 1040 10451050 aat atc ttg cac aag caa ctg ctg aac aat atc ctt cga gca cct 3204Asn Ile Leu His Lys Gln Leu Leu Asn Asn Ile Leu Arg Ala Pro 1055 10601065 atg aga ttt ttt gac aca aca ccc aca ggc cgg att gtg aac agg 3249Met Arg Phe Phe Asp Thr Thr Pro Thr Gly Arg Ile Val Asn Arg 1070 10751080 ttt gcc ggc gat att tcc aca gtg gat gac acc ctg cct cag tcc 3294Phe Ala Gly Asp Ile Ser Thr Val Asp Asp Thr Leu Pro Gln Ser 1085 10901095 ttg cgc agc tgg att aca tgc ttc ctg ggg ata atc agc acc ctt 3339Leu Arg Ser Trp Ile Thr Cys Phe Leu Gly Ile Ile Ser Thr Leu 1100 11051110 gtc atg atc tgc atg gcc act cct gtc ttc acc atc atc gtc att 3384Val Met Ile Cys Met Ala Thr Pro Val Phe Thr Ile Ile Val Ile 1115 11201125 cct ctt ggc att att tat gta tct gtt cag atg ttt tat gtg tct 3429Pro Leu Gly Ile Ile Tyr Val Ser Val Gln Met Phe Tyr Val Ser 1130 11351140 acc tcc cgc cag ctg agg cgt ctg gac tct gtc acc agg tcc cca 3474Thr Ser Arg Gln Leu Arg Arg Leu Asp Ser Val Thr Arg Ser Pro 1145 11501155 atc tac tct cac ttc agc gag acc gta tca ggt ttg cca gtt atc 3519Ile Tyr Ser His Phe Ser Glu Thr Val Ser Gly Leu Pro Val Ile 1160 11651170 cgt gcc ttt gag cac cag cag cga ttt ctg aaa cac aat gag gag 3564Arg Ala Phe Glu His Gln Gln Arg Phe Leu Lys His Asn Glu Glu 1175 11801185 agg att gac acc aac cag aaa tgt gtc ttt tcc tgg atc acc tcc 3609Arg Ile Asp Thr Asn Gln Lys Cys Val Phe Ser Trp Ile Thr Ser 1190 11951200 aac agg tgg ctt gca att cgc ctg gag ctg gtt ggg aac ctg act 3654Asn Arg Trp Leu Ala Ile Arg Leu Glu Leu Val Gly Asn Leu Thr 1205 12101215 gtc ttc ttt tca gcc ttg atg atg gtt att tat aga gat acc cta 3699Val Phe Phe Ser Ala Leu Met Met Val Ile Tyr Arg Asp Thr Leu 1220 12251230 agt ggg gac act gtt ggc ttt gtt ctg tcc aat gca ctc aat atc 3744Ser Gly Asp Thr Val Gly Phe Val Leu Ser Asn Ala Leu Asn Ile 1235 12401245 aca caa acc ctg aac tgg ctg gtg agg atg aca tca gaa ata gag 3789Thr Gln Thr Leu Asn Trp Leu Val Arg Met Thr Ser Glu Ile Glu 1250 12551260 acc aac att gtg gct gtt gag cga ata act gag tac aca aaa gtg 3834Thr Asn Ile Val Ala Val Glu Arg Ile Thr Glu Tyr Thr Lys Val 1265 12701275 gaa aat gag gca ccc tgg gtg act gat aag agg cct ccg cca gat 3879Glu Asn Glu Ala Pro Trp Val Thr Asp Lys Arg Pro Pro Pro Asp 1280 12851290 tgg ccc agc aaa ggc aag atc cag ttt aac aac tac caa gtg cgg 3924Trp Pro Ser Lys Gly Lys Ile Gln Phe Asn Asn Tyr Gln Val Arg 1295 13001305 tac cga cct gag ctg gat ctg gtc ctc aga ggg atc act tgt gac 3969Tyr Arg Pro Glu Leu Asp Leu Val Leu Arg Gly Ile Thr Cys Asp 1310 13151320 atc ggt agc atg gag aag att ggt gtg gtg ggc agg aca gga gct 4014Ile Gly Ser Met Glu Lys Ile Gly Val Val Gly Arg Thr Gly Ala 1325 13301335 gga aag tca tcc ctc aca aac tgc ctc ttc aga atc tta gag gct 4059Gly Lys Ser Ser Leu Thr Asn Cys Leu Phe Arg Ile Leu Glu Ala 1340 13451350 gcc ggt ggt cag att atc att gat gga gta gat att gct tcc att 4104Ala Gly Gly Gln Ile Ile Ile Asp Gly Val Asp Ile Ala Ser Ile 1355 13601365 ggg ctc cac gac ctc cga gag aag ctg acc atc atc ccc cag gac 4149Gly Leu His Asp Leu Arg Glu Lys Leu Thr Ile Ile Pro Gln Asp 1370 13751380 ccc atc ctg ttc tct gga agc ctg agg atg aat ctc gac cct ttc 4194Pro Ile Leu Phe Ser Gly Ser Leu Arg Met Asn Leu Asp Pro Phe 1385 13901395 aac aac tac tca gat gag gag att tgg aag gcc ttg gag ctg gct 4239Asn Asn Tyr Ser Asp Glu Glu Ile Trp Lys Ala Leu Glu Leu Ala 1400 14051410 cac ctc aag tct ttt gtg gcc agc ctg caa ctt ggg tta tcc cac 4284His Leu Lys Ser Phe Val Ala Ser Leu Gln Leu Gly Leu Ser His 1415 14201425 gaa gtt aca gag gct ggt ggc aac ctg agc ata ggc cag agg cag 4329Glu Val Thr Glu Ala Gly Gly Asn Leu Ser Ile Gly Gln Arg Gln 1430 14351440 ctg ctg tgc ctg ggc agg gct ctg ctt cgg aaa tcc aag atc ctg 4374Leu Leu Cys Leu Gly Arg Ala Leu Leu Arg Lys Ser Lys Ile Leu 1445 14501455 gtc ctg gat gag gcc act gct gcg gtg gat cta gag aca gac aac 4419Val Leu Asp Glu Ala Thr Ala Ala Val Asp Leu Glu Thr Asp Asn 1460 14651470 ctc att cag acg acc atc caa aac gag ttc gcc cac tgc aca gtg 4464Leu Ile Gln Thr Thr Ile Gln Asn Glu Phe Ala His Cys Thr Val 1475 14801485 atc acc atc gcc cac agg ctg cat acc atc atg gac agt gac aag 4509Ile Thr Ile Ala His Arg Leu His Thr Ile Met Asp Ser Asp Lys 1490 14951500 gta atg gtc cta gac aac ggg aag att ata gag tac ggc agc cct 4554Val Met Val Leu Asp Asn Gly Lys Ile Ile Glu Tyr Gly Ser Pro 1505 15101515 gaa gaa ctg cta caa atc cct gga ccc ttt tac ttt atg gct aag 4599Glu Glu Leu Leu Gln Ile Pro Gly Pro Phe Tyr Phe Met Ala Lys 1520 15251530 gaa gct ggc att gag aat gtg aac agc aca gca ttc gcaccg 4641 Glu AlaGly Ile Glu Asn Val Asn Ser Thr Ala Phe 1535 1540 1545 12 1545 PRT Homosapiens 12 Met Leu Glu Lys Phe Cys Asn Ser Thr Phe Trp Asn Ser Ser PheLeu 1 5 10 15 Asp Ser Pro Glu Ala Asp Leu Pro Leu Cys Phe Glu Gln ThrVal Leu 20 25 30 Val Trp Ile Pro Leu Gly Phe Leu Trp Leu Leu Ala Pro TrpGln Leu 35 40 45 Leu His Val Tyr Lys Ser Arg Thr Lys Arg Ser Ser Thr ThrLys Leu 50 55 60 Tyr Leu Ala Lys Gln Val Phe Val Gly Phe Leu Leu Ile LeuAla Ala 65 70 75 80 Ile Glu Leu Ala Leu Val Leu Thr Glu Asp Ser Gly GlnAla Thr Val 85 90 95 Pro Ala Val Arg Tyr Thr Asn Pro Ser Leu Tyr Leu GlyThr Trp Leu 100 105 110 Leu Val Leu Leu Ile Gln Tyr Ser Arg Gln Trp CysVal Gln Lys Asn 115 120 125 Ser Trp Phe Leu Ser Leu Phe Trp Ile Leu SerIle Leu Cys Gly Thr 130 135 140 Phe Gln Phe Gln Thr Leu Ile Arg Thr LeuLeu Gln Gly Asp Asn Ser 145 150 155 160 Asn Leu Ala Tyr Ser Cys Leu PhePhe Ile Ser Tyr Gly Phe Gln Ile 165 170 175 Leu Ile Leu Ile Phe Ser AlaPhe Ser Glu Asn Asn Glu Ser Ser Asn 180 185 190 Asn Pro Ser Ser Ile AlaSer Phe Leu Ser Ser Ile Thr Tyr Ser Trp 195 200 205 Tyr Asp Ser Ile IleLeu Lys Gly Tyr Lys Arg Pro Leu Thr Leu Glu 210 215 220 Asp Val Trp GluVal Asp Glu Glu Met Lys Thr Lys Thr Leu Val Ser 225 230 235 240 Lys PheGlu Thr His Met Lys Arg Glu Leu Gln Lys Ala Arg Arg Ala 245 250 255 LeuGln Arg Arg Gln Glu Lys Ser Ser Gln Gln Asn Ser Gly Ala Arg 260 265 270Leu Pro Gly Leu Asn Lys Asn Gln Ser Gln Ser Gln Asp Ala Leu Val 275 280285 Leu Glu Asp Val Glu Lys Lys Lys Lys Lys Ser Gly Thr Lys Lys Asp 290295 300 Val Pro Lys Ser Trp Leu Met Lys Ala Leu Phe Lys Thr Phe Tyr Met305 310 315 320 Val Leu Leu Lys Ser Phe Leu Leu Lys Leu Val Asn Asp IlePhe Thr 325 330 335 Phe Val Ser Pro Gln Leu Leu Lys Leu Leu Ile Ser PheAla Ser Asp 340 345 350 Arg Asp Thr Tyr Leu Trp Ile Gly Tyr Leu Cys AlaIle Leu Leu Phe 355 360 365 Thr Ala Ala Leu Ile Gln Ser Phe Cys Leu GlnCys Tyr Phe Gln Leu 370 375 380 Cys Phe Lys Leu Gly Val Lys Val Arg ThrAla Ile Met Ala Ser Val 385 390 395 400 Tyr Lys Lys Ala Leu Thr Leu SerAsn Leu Ala Arg Lys Glu Tyr Thr 405 410 415 Val Gly Glu Thr Val Asn LeuMet Ser Val Asp Ala Gln Lys Leu Met 420 425 430 Asp Val Thr Asn Phe MetHis Met Leu Trp Ser Ser Val Leu Gln Ile 435 440 445 Val Leu Ser Ile PhePhe Leu Trp Arg Glu Leu Gly Pro Ser Val Leu 450 455 460 Ala Gly Val GlyVal Met Val Leu Val Ile Pro Ile Asn Ala Ile Leu 465 470 475 480 Ser ThrLys Ser Lys Thr Ile Gln Val Lys Asn Met Lys Asn Lys Asp 485 490 495 LysArg Leu Lys Ile Met Asn Glu Ile Leu Ser Gly Ile Lys Ile Leu 500 505 510Lys Tyr Phe Ala Trp Glu Pro Ser Phe Arg Asp Gln Val Gln Asn Leu 515 520525 Arg Lys Lys Glu Leu Lys Asn Leu Leu Ala Phe Ser Gln Leu Gln Cys 530535 540 Val Val Ile Phe Val Phe Gln Leu Thr Pro Val Leu Val Ser Val Val545 550 555 560 Thr Phe Ser Val Tyr Val Leu Val Asp Ser Asn Asn Ile LeuAsp Ala 565 570 575 Gln Lys Ala Phe Thr Ser Ile Thr Leu Phe Asn Ile LeuArg Phe Pro 580 585 590 Leu Ser Met Leu Pro Met Met Ile Ser Ser Met LeuGln Ala Ser Val 595 600 605 Ser Thr Glu Arg Leu Glu Lys Tyr Leu Gly GlyAsp Asp Leu Asp Thr 610 615 620 Ser Ala Ile Arg His Asp Cys Asn Phe AspLys Ala Met Gln Phe Ser 625 630 635 640 Glu Ala Ser Phe Thr Trp Glu HisAsp Ser Glu Ala Thr Val Arg Asp 645 650 655 Val Asn Leu Asp Ile Met AlaGly Gln Leu Val Ala Val Ile Gly Pro 660 665 670 Val Gly Ser Gly Lys SerSer Leu Ile Ser Ala Met Leu Gly Glu Met 675 680 685 Glu Asn Val His GlyHis Ile Thr Ile Lys Gly Thr Thr Ala Tyr Val 690 695 700 Pro Gln Gln SerTrp Ile Gln Asn Gly Thr Ile Lys Asp Asn Ile Leu 705 710 715 720 Phe GlyThr Glu Phe Asn Glu Lys Arg Tyr Gln Gln Val Leu Glu Ala 725 730 735 CysAla Leu Leu Pro Asp Leu Glu Met Leu Pro Gly Gly Asp Leu Ala 740 745 750Glu Ile Gly Glu Lys Gly Ile Asn Leu Ser Gly Gly Gln Lys Gln Arg 755 760765 Ile Ser Leu Ala Arg Ala Thr Tyr Gln Asn Leu Asp Ile Tyr Leu Leu 770775 780 Asp Asp Pro Leu Ser Ala Val Asp Ala His Val Gly Lys His Ile Phe785 790 795 800 Asn Lys Val Leu Gly Pro Asn Gly Leu Leu Lys Gly Lys ThrArg Leu 805 810 815 Leu Val Thr His Ser Met His Phe Leu Pro Gln Val AspGlu Ile Val 820 825 830 Val Leu Gly Asn Gly Thr Ile Val Glu Lys Gly SerTyr Ser Ala Leu 835 840 845 Leu Ala Lys Lys Gly Glu Phe Ala Lys Asn LeuLys Thr Phe Leu Arg 850 855 860 His Thr Gly Pro Glu Glu Glu Ala Thr ValHis Asp Gly Ser Glu Glu 865 870 875 880 Glu Asp Asp Asp Tyr Gly Leu IleSer Ser Val Glu Glu Ile Pro Glu 885 890 895 Asp Ala Ala Ser Ile Thr MetArg Arg Glu Asn Ser Phe Arg Arg Thr 900 905 910 Leu Ser Arg Ser Ser ArgSer Asn Gly Arg His Leu Lys Ser Leu Arg 915 920 925 Asn Ser Leu Lys ThrArg Asn Val Asn Ser Leu Lys Glu Asp Glu Glu 930 935 940 Leu Val Lys GlyGln Lys Leu Ile Lys Lys Glu Phe Ile Glu Thr Gly 945 950 955 960 Lys ValLys Phe Ser Ile Tyr Leu Glu Tyr Leu Gln Ala Ile Gly Leu 965 970 975 PheSer Ile Phe Phe Ile Ile Leu Ala Phe Val Met Asn Ser Val Ala 980 985 990Phe Ile Gly Ser Asn Leu Trp Leu Ser Ala Trp Thr Ser Asp Ser Lys 995 10001005 Ile Phe Asn Ser Thr Asp Tyr Pro Ala Ser Gln Arg Asp Met Arg 10101015 1020 Val Gly Val Tyr Gly Ala Leu Gly Leu Ala Gln Gly Ile Phe Val1025 1030 1035 Phe Ile Ala His Phe Trp Ser Ala Phe Gly Phe Val His AlaSer 1040 1045 1050 Asn Ile Leu His Lys Gln Leu Leu Asn Asn Ile Leu ArgAla Pro 1055 1060 1065 Met Arg Phe Phe Asp Thr Thr Pro Thr Gly Arg IleVal Asn Arg 1070 1075 1080 Phe Ala Gly Asp Ile Ser Thr Val Asp Asp ThrLeu Pro Gln Ser 1085 1090 1095 Leu Arg Ser Trp Ile Thr Cys Phe Leu GlyIle Ile Ser Thr Leu 1100 1105 1110 Val Met Ile Cys Met Ala Thr Pro ValPhe Thr Ile Ile Val Ile 1115 1120 1125 Pro Leu Gly Ile Ile Tyr Val SerVal Gln Met Phe Tyr Val Ser 1130 1135 1140 Thr Ser Arg Gln Leu Arg ArgLeu Asp Ser Val Thr Arg Ser Pro 1145 1150 1155 Ile Tyr Ser His Phe SerGlu Thr Val Ser Gly Leu Pro Val Ile 1160 1165 1170 Arg Ala Phe Glu HisGln Gln Arg Phe Leu Lys His Asn Glu Glu 1175 1180 1185 Arg Ile Asp ThrAsn Gln Lys Cys Val Phe Ser Trp Ile Thr Ser 1190 1195 1200 Asn Arg TrpLeu Ala Ile Arg Leu Glu Leu Val Gly Asn Leu Thr 1205 1210 1215 Val PhePhe Ser Ala Leu Met Met Val Ile Tyr Arg Asp Thr Leu 1220 1225 1230 SerGly Asp Thr Val Gly Phe Val Leu Ser Asn Ala Leu Asn Ile 1235 1240 1245Thr Gln Thr Leu Asn Trp Leu Val Arg Met Thr Ser Glu Ile Glu 1250 12551260 Thr Asn Ile Val Ala Val Glu Arg Ile Thr Glu Tyr Thr Lys Val 12651270 1275 Glu Asn Glu Ala Pro Trp Val Thr Asp Lys Arg Pro Pro Pro Asp1280 1285 1290 Trp Pro Ser Lys Gly Lys Ile Gln Phe Asn Asn Tyr Gln ValArg 1295 1300 1305 Tyr Arg Pro Glu Leu Asp Leu Val Leu Arg Gly Ile ThrCys Asp 1310 1315 1320 Ile Gly Ser Met Glu Lys Ile Gly Val Val Gly ArgThr Gly Ala 1325 1330 1335 Gly Lys Ser Ser Leu Thr Asn Cys Leu Phe ArgIle Leu Glu Ala 1340 1345 1350 Ala Gly Gly Gln Ile Ile Ile Asp Gly ValAsp Ile Ala Ser Ile 1355 1360 1365 Gly Leu His Asp Leu Arg Glu Lys LeuThr Ile Ile Pro Gln Asp 1370 1375 1380 Pro Ile Leu Phe Ser Gly Ser LeuArg Met Asn Leu Asp Pro Phe 1385 1390 1395 Asn Asn Tyr Ser Asp Glu GluIle Trp Lys Ala Leu Glu Leu Ala 1400 1405 1410 His Leu Lys Ser Phe ValAla Ser Leu Gln Leu Gly Leu Ser His 1415 1420 1425 Glu Val Thr Glu AlaGly Gly Asn Leu Ser Ile Gly Gln Arg Gln 1430 1435 1440 Leu Leu Cys LeuGly Arg Ala Leu Leu Arg Lys Ser Lys Ile Leu 1445 1450 1455 Val Leu AspGlu Ala Thr Ala Ala Val Asp Leu Glu Thr Asp Asn 1460 1465 1470 Leu IleGln Thr Thr Ile Gln Asn Glu Phe Ala His Cys Thr Val 1475 1480 1485 IleThr Ile Ala His Arg Leu His Thr Ile Met Asp Ser Asp Lys 1490 1495 1500Val Met Val Leu Asp Asn Gly Lys Ile Ile Glu Tyr Gly Ser Pro 1505 15101515 Glu Glu Leu Leu Gln Ile Pro Gly Pro Phe Tyr Phe Met Ala Lys 15201525 1530 Glu Ala Gly Ile Glu Asn Val Asn Ser Thr Ala Phe 1535 1540 154513 4645 DNA Homo sapiens CDS (1)..(4635) 13 atg ctg gag aag ttc tgc aactct act ttt tgg aat tcc tca ttc ctg 48 Met Leu Glu Lys Phe Cys Asn SerThr Phe Trp Asn Ser Ser Phe Leu 1 5 10 15 gac agt ccg gag gca gac ctgcca ctt tgt ttt gag caa act gtt ctg 96 Asp Ser Pro Glu Ala Asp Leu ProLeu Cys Phe Glu Gln Thr Val Leu 20 25 30 gtg tgg att ccc ttg ggc ttc ctatgg ctc ctg gcc ccc tgg cag ctt 144 Val Trp Ile Pro Leu Gly Phe Leu TrpLeu Leu Ala Pro Trp Gln Leu 35 40 45 ctc cac gtg tat aaa tcc agg acc aagaga tcc tct acc acc aaa ctc 192 Leu His Val Tyr Lys Ser Arg Thr Lys ArgSer Ser Thr Thr Lys Leu 50 55 60 tat ctt gct aag cag gta ttc gtt ggt tttctt ctt att cta gca gcc 240 Tyr Leu Ala Lys Gln Val Phe Val Gly Phe LeuLeu Ile Leu Ala Ala 65 70 75 80 ata gag ctg gcc ctt gta ctc aca gaa gactct gga caa gcc aca gtc 288 Ile Glu Leu Ala Leu Val Leu Thr Glu Asp SerGly Gln Ala Thr Val 85 90 95 cct gct gtt cga tat acc aat cca agc ctc taccta ggc aca tgg ctc 336 Pro Ala Val Arg Tyr Thr Asn Pro Ser Leu Tyr LeuGly Thr Trp Leu 100 105 110 ctg gtt ttg ctg atc caa tac agc aga caa tggtgt gta cag aaa aac 384 Leu Val Leu Leu Ile Gln Tyr Ser Arg Gln Trp CysVal Gln Lys Asn 115 120 125 tcc tgg ttc ctg tcc cta ttc tgg att ctc tcgata ctc tgt ggc act 432 Ser Trp Phe Leu Ser Leu Phe Trp Ile Leu Ser IleLeu Cys Gly Thr 130 135 140 ttc caa ttt cag act ctg atc cgg aca ctc ttacag ggt gac aat tct 480 Phe Gln Phe Gln Thr Leu Ile Arg Thr Leu Leu GlnGly Asp Asn Ser 145 150 155 160 aat cta gcc tac tcc tgc ctg ttc ttc atctcc tac gga ttc cag atc 528 Asn Leu Ala Tyr Ser Cys Leu Phe Phe Ile SerTyr Gly Phe Gln Ile 165 170 175 ctg atc ctg atc ttt tca gca ttt tca gaaaat aat gag tca tca aat 576 Leu Ile Leu Ile Phe Ser Ala Phe Ser Glu AsnAsn Glu Ser Ser Asn 180 185 190 aat cca tca tcc ata gct tca ttc ctg agtagc att acc tac agc tgg 624 Asn Pro Ser Ser Ile Ala Ser Phe Leu Ser SerIle Thr Tyr Ser Trp 195 200 205 tat gac agc atc att ctg aaa ggc tac aagcgt cct ctg aca ctc gag 672 Tyr Asp Ser Ile Ile Leu Lys Gly Tyr Lys ArgPro Leu Thr Leu Glu 210 215 220 gat gtc tgg gaa gtt gat gaa gag atg aaaacc aag aca tta gtg agc 720 Asp Val Trp Glu Val Asp Glu Glu Met Lys ThrLys Thr Leu Val Ser 225 230 235 240 aag ttt gaa acg cac atg aag aga gagctg cag aaa gcc agg cgg gca 768 Lys Phe Glu Thr His Met Lys Arg Glu LeuGln Lys Ala Arg Arg Ala 245 250 255 ctc cag aga cgg cag gag aag agc tcccag cag aac tct gga gcc agg 816 Leu Gln Arg Arg Gln Glu Lys Ser Ser GlnGln Asn Ser Gly Ala Arg 260 265 270 ctg cct ggc ttg aac aag aat cag agtcaa agc caa gat gcc ctt gtc 864 Leu Pro Gly Leu Asn Lys Asn Gln Ser GlnSer Gln Asp Ala Leu Val 275 280 285 ctg gaa gat gtt gaa aag aaa aaa aagaag tct ggg acc aaa aaa gat 912 Leu Glu Asp Val Glu Lys Lys Lys Lys LysSer Gly Thr Lys Lys Asp 290 295 300 gtt cca aaa tcc tgg ttg atg aag gctctg ttc aaa act ttc tac atg 960 Val Pro Lys Ser Trp Leu Met Lys Ala LeuPhe Lys Thr Phe Tyr Met 305 310 315 320 gtg ctc ctg aaa tca ttc cta ctgaag cta gtg aat gac atc ttc acg 1008 Val Leu Leu Lys Ser Phe Leu Leu LysLeu Val Asn Asp Ile Phe Thr 325 330 335 ttt gtg agt cct cag ctg ctg aaattg ctg atc tcc ttt gca agt gac 1056 Phe Val Ser Pro Gln Leu Leu Lys LeuLeu Ile Ser Phe Ala Ser Asp 340 345 350 cgt gac aca tat ttg tgg att ggatat ctc tgt gca atc ctc tta ttc 1104 Arg Asp Thr Tyr Leu Trp Ile Gly TyrLeu Cys Ala Ile Leu Leu Phe 355 360 365 act gcg gct ctc att cag tct ttctgc ctt cag tgt tat ttc caa ctg 1152 Thr Ala Ala Leu Ile Gln Ser Phe CysLeu Gln Cys Tyr Phe Gln Leu 370 375 380 tgc ttc aag ctg ggt gta aaa gtacgg aca gct atc atg gct tct gta 1200 Cys Phe Lys Leu Gly Val Lys Val ArgThr Ala Ile Met Ala Ser Val 385 390 395 400 tat aag aag gca ttg acc ctatcc aac ttg gcc agg aag gag tac acc 1248 Tyr Lys Lys Ala Leu Thr Leu SerAsn Leu Ala Arg Lys Glu Tyr Thr 405 410 415 gtt gga gaa aca gtg aac ctgatg tct gtg gat gcc cag aag ctc atg 1296 Val Gly Glu Thr Val Asn Leu MetSer Val Asp Ala Gln Lys Leu Met 420 425 430 gat gtg acc aac ttc atg cacatg ctg tgg tca agt gtt cta cag att 1344 Asp Val Thr Asn Phe Met His MetLeu Trp Ser Ser Val Leu Gln Ile 435 440 445 gtc tta tct atc ttc ttc ctatgg aga gag ttg gga ccc tca gtc tta 1392 Val Leu Ser Ile Phe Phe Leu TrpArg Glu Leu Gly Pro Ser Val Leu 450 455 460 gca ggt gtt ggg gtg atg gtgctt gta atc cca att aat gcg ata ctg 1440 Ala Gly Val Gly Val Met Val LeuVal Ile Pro Ile Asn Ala Ile Leu 465 470 475 480 tcc acc aag agt aag accatt cag gtc aaa aat atg aag aat aaa gac 1488 Ser Thr Lys Ser Lys Thr IleGln Val Lys Asn Met Lys Asn Lys Asp 485 490 495 aaa cgt tta aag atc atgaat gag att ctt agt gga atc aag atc ctg 1536 Lys Arg Leu Lys Ile Met AsnGlu Ile Leu Ser Gly Ile Lys Ile Leu 500 505 510 aaa tat ttt gcc tgg gaacct tca ttc aga gac caa gta caa aac ctc 1584 Lys Tyr Phe Ala Trp Glu ProSer Phe Arg Asp Gln Val Gln Asn Leu 515 520 525 cgg aag aaa gag ctc aagaac ctg ctg gcc ttt agt caa cta cag tgt 1632 Arg Lys Lys Glu Leu Lys AsnLeu Leu Ala Phe Ser Gln Leu Gln Cys 530 535 540 gta gta ata ttc gtc ttccag tta act cca gtc ctg gta tct gtg gtc 1680 Val Val Ile Phe Val Phe GlnLeu Thr Pro Val Leu Val Ser Val Val 545 550 555 560 aca ttt tct gtt tatgtc ctg gtg gat agc aac aat att ttg gat gca 1728 Thr Phe Ser Val Tyr ValLeu Val Asp Ser Asn Asn Ile Leu Asp Ala 565 570 575 caa aag gcc ttc acctcc att acc ctc ttc aat atc ctg cgc ttt ccc 1776 Gln Lys Ala Phe Thr SerIle Thr Leu Phe Asn Ile Leu Arg Phe Pro 580 585 590 ctg agc atg ctt cccatg atg atc tcc tcc atg ctc cag gcc agt gtt 1824 Leu Ser Met Leu Pro MetMet Ile Ser Ser Met Leu Gln Ala Ser Val 595 600 605 tcc aca gag cgg ctagag aag tac ttg gga ggg gat gac ttg gac aca 1872 Ser Thr Glu Arg Leu GluLys Tyr Leu Gly Gly Asp Asp Leu Asp Thr 610 615 620 tct gcc att cga catgac tgc aat ttt gac aaa gcc atg cag ttt tct 1920 Ser Ala Ile Arg His AspCys Asn Phe Asp Lys Ala Met Gln Phe Ser 625 630 635 640 gag gcc tcc tttacc tgg gaa cat gat tcg gaa gcc aca gtc cga gat 1968 Glu Ala Ser Phe ThrTrp Glu His Asp Ser Glu Ala Thr Val Arg Asp 645 650 655 gtg aac ctg gacatt atg gca ggc caa ctt gtg gct gtg ata ggc cct 2016 Val Asn Leu Asp IleMet Ala Gly Gln Leu Val Ala Val Ile Gly Pro 660 665 670 gtc ggc tct gggaaa tcc tcc ttg ata tca gcc atg ctg gga gaa atg 2064 Val Gly Ser Gly LysSer Ser Leu Ile Ser Ala Met Leu Gly Glu Met 675 680 685 gaa aat gtc cacggg cac atc acc atc aag ggc acc act gcc tat gtc 2112 Glu Asn Val His GlyHis Ile Thr Ile Lys Gly Thr Thr Ala Tyr Val 690 695 700 cca cag cag tcctgg att cag aat ggc acc ata aag gac aac atc ctt 2160 Pro Gln Gln Ser TrpIle Gln Asn Gly Thr Ile Lys Asp Asn Ile Leu 705 710 715 720 ttt gga acagag ttt aat gaa aag agg tac cag caa gta ctg gag gcc 2208 Phe Gly Thr GluPhe Asn Glu Lys Arg Tyr Gln Gln Val Leu Glu Ala 725 730 735 tgt gct ctcctc cca gac ttg gaa atg ctg cct gga gga gat ttg gct 2256 Cys Ala Leu LeuPro Asp Leu Glu Met Leu Pro Gly Gly Asp Leu Ala 740 745 750 gag att ggagag aag ggt ata aat ctt agt ggg ggt cag aag cag cgg 2304 Glu Ile Gly GluLys Gly Ile Asn Leu Ser Gly Gly Gln Lys Gln Arg 755 760 765 atc agc ctggcc aga gct acc tac caa aat tta gac atc tat ctt cta 2352 Ile Ser Leu AlaArg Ala Thr Tyr Gln Asn Leu Asp Ile Tyr Leu Leu 770 775 780 gat gac cccctg tct gca gtg gat gct cat gta gga aaa cat att ttt 2400 Asp Asp Pro LeuSer Ala Val Asp Ala His Val Gly Lys His Ile Phe 785 790 795 800 aat aaggtc ttg ggc ccc aat ggc ctg ttg aaa ggc aag act cga ctc 2448 Asn Lys ValLeu Gly Pro Asn Gly Leu Leu Lys Gly Lys Thr Arg Leu 805 810 815 ttg gttaca cat agc atg cac ttt ctt cct caa gtg gat gag att gta 2496 Leu Val ThrHis Ser Met His Phe Leu Pro Gln Val Asp Glu Ile Val 820 825 830 gtt ctgggg aat gga aca att gta gag aaa gga tcc tac agt gct ctc 2544 Val Leu GlyAsn Gly Thr Ile Val Glu Lys Gly Ser Tyr Ser Ala Leu 835 840 845 ctg gccaaa aaa gga gag ttt gct aag aat ctg aag aca ttt cta aga 2592 Leu Ala LysLys Gly Glu Phe Ala Lys Asn Leu Lys Thr Phe Leu Arg 850 855 860 cat acaggc cct gaa gag gaa gcc aca gtc cat gat ggc agt gaa gaa 2640 His Thr GlyPro Glu Glu Glu Ala Thr Val His Asp Gly Ser Glu Glu 865 870 875 880 gaagac gat gac tat ggg ctg ata tcc agt gtg gaa gag atc ccc gaa 2688 Glu AspAsp Asp Tyr Gly Leu Ile Ser Ser Val Glu Glu Ile Pro Glu 885 890 895 gatgca gcc tcc ata acc atg aga aga gag aac agc ttt cgt cga aca 2736 Asp AlaAla Ser Ile Thr Met Arg Arg Glu Asn Ser Phe Arg Arg Thr 900 905 910 cttagc cgc agt tct agg tcc aat ggc agg cat ctg aag tcc ctg aga 2784 Leu SerArg Ser Ser Arg Ser Asn Gly Arg His Leu Lys Ser Leu Arg 915 920 925 aactcc ttg aaa act cgg aat gtg aat agc ctg aag gaa gac gaa gaa 2832 Asn SerLeu Lys Thr Arg Asn Val Asn Ser Leu Lys Glu Asp Glu Glu 930 935 940 ctagtg aaa gga caa aaa cta att aag aag gaa ttc ata gaa act gga 2880 Leu ValLys Gly Gln Lys Leu Ile Lys Lys Glu Phe Ile Glu Thr Gly 945 950 955 960aag gtg aag ttc tcc atc tac ctg gag tac cta caa gca ata gga ttg 2928 LysVal Lys Phe Ser Ile Tyr Leu Glu Tyr Leu Gln Ala Ile Gly Leu 965 970 975ttt tcg ata ttc ttc atc atc ctt gcg ttt gtg atg aat tct gtg gct 2976 PheSer Ile Phe Phe Ile Ile Leu Ala Phe Val Met Asn Ser Val Ala 980 985 990ttt att gga tcc aac ctc tgg ctc agt gct tgg acc agt gac tct aaa 3024 PheIle Gly Ser Asn Leu Trp Leu Ser Ala Trp Thr Ser Asp Ser Lys 995 10001005 atc ttc aat agc acc gac tat cca gca tct cag agg gac atg aga 3069Ile Phe Asn Ser Thr Asp Tyr Pro Ala Ser Gln Arg Asp Met Arg 1010 10151020 gtt gga gtc tac gga gct ctg gga tta gcc caa ggt ata ttt gtg 3114Val Gly Val Tyr Gly Ala Leu Gly Leu Ala Gln Gly Ile Phe Val 1025 10301035 ttc ata gca cat ttc tgg agt gcc ttt ggt ttc gtc cat gca tca 3159Phe Ile Ala His Phe Trp Ser Ala Phe Gly Phe Val His Ala Ser 1040 10451050 aat atc ttg cac aag caa ctg ctg aac aat atc ctt cga gca cct 3204Asn Ile Leu His Lys Gln Leu Leu Asn Asn Ile Leu Arg Ala Pro 1055 10601065 atg aga ttt ttt gac aca aca ccc aca ggc cgg att gtg aac agg 3249Met Arg Phe Phe Asp Thr Thr Pro Thr Gly Arg Ile Val Asn Arg 1070 10751080 ttt gcc ggc gat att tcc aca gtg gat gac acc ctg cct cag tcc 3294Phe Ala Gly Asp Ile Ser Thr Val Asp Asp Thr Leu Pro Gln Ser 1085 10901095 ttg cgc agc tgg att aca tgc ttc ctg ggg ata atc agc acc ctt 3339Leu Arg Ser Trp Ile Thr Cys Phe Leu Gly Ile Ile Ser Thr Leu 1100 11051110 gtc atg atc tgc atg gcc act cct gtc ttc acc atc atc gtc att 3384Val Met Ile Cys Met Ala Thr Pro Val Phe Thr Ile Ile Val Ile 1115 11201125 cct ctt ggc att att tat gta tct gtt cag atg ttt tat gtg tct 3429Pro Leu Gly Ile Ile Tyr Val Ser Val Gln Met Phe Tyr Val Ser 1130 11351140 acc tcc cgc cag ctg agg cgt ctg gac tct gtc acc agg tcc cca 3474Thr Ser Arg Gln Leu Arg Arg Leu Asp Ser Val Thr Arg Ser Pro 1145 11501155 atc tac tct cac ttc agc gag acc gta tca ggt ttg cca gtt atc 3519Ile Tyr Ser His Phe Ser Glu Thr Val Ser Gly Leu Pro Val Ile 1160 11651170 cgt gcc ttt gag cac cag cag cga ttt ctg aaa cac aat gag gag 3564Arg Ala Phe Glu His Gln Gln Arg Phe Leu Lys His Asn Glu Glu 1175 11801185 agg att gac acc aac cag aaa tgt gtc ttt tcc tgg atc acc tcc 3609Arg Ile Asp Thr Asn Gln Lys Cys Val Phe Ser Trp Ile Thr Ser 1190 11951200 aac agg tgg ctt gca att cgc ctg gag ctg gtt ggg aac ctg act 3654Asn Arg Trp Leu Ala Ile Arg Leu Glu Leu Val Gly Asn Leu Thr 1205 12101215 gtc ttc ttt tca gcc ttg atg atg gtt att tat aga gat acc cta 3699Val Phe Phe Ser Ala Leu Met Met Val Ile Tyr Arg Asp Thr Leu 1220 12251230 agt ggg gac act gtt ggc ttt gtt ctg tcc aat gca ctc aat atc 3744Ser Gly Asp Thr Val Gly Phe Val Leu Ser Asn Ala Leu Asn Ile 1235 12401245 aca caa acc ctg aac tgg ctg gtg agg atg aca tca gaa ata gag 3789Thr Gln Thr Leu Asn Trp Leu Val Arg Met Thr Ser Glu Ile Glu 1250 12551260 acc aac att gtg gct gtt gag cga ata act gag tac aca aaa gtg 3834Thr Asn Ile Val Ala Val Glu Arg Ile Thr Glu Tyr Thr Lys Val 1265 12701275 gaa aat gag gca ccc tgg gtg act gat aag agg cct ccg cca gat 3879Glu Asn Glu Ala Pro Trp Val Thr Asp Lys Arg Pro Pro Pro Asp 1280 12851290 tgg ccc agc aaa ggc aag atc cag ttt aac aac tac caa gtg cgg 3924Trp Pro Ser Lys Gly Lys Ile Gln Phe Asn Asn Tyr Gln Val Arg 1295 13001305 tac cga cct gag ctg gat ctg gtc ctc aga ggg atc act tgt gac 3969Tyr Arg Pro Glu Leu Asp Leu Val Leu Arg Gly Ile Thr Cys Asp 1310 13151320 atc ggt agc atg gag aag att ggt gtg gtg ggc agg aca gga gct 4014Ile Gly Ser Met Glu Lys Ile Gly Val Val Gly Arg Thr Gly Ala 1325 13301335 gga aag tca tcc ctc aca aac tgc ctc ttc aga atc tta gag gct 4059Gly Lys Ser Ser Leu Thr Asn Cys Leu Phe Arg Ile Leu Glu Ala 1340 13451350 gcc ggt ggt cag att atc att gat gga gta gat att gct tcc att 4104Ala Gly Gly Gln Ile Ile Ile Asp Gly Val Asp Ile Ala Ser Ile 1355 13601365 ggg ctc cac gac ctc cga gag aag ctg acc atc atc ccc cag gac 4149Gly Leu His Asp Leu Arg Glu Lys Leu Thr Ile Ile Pro Gln Asp 1370 13751380 ccc atc ctg ttc tct gga agc ctg agg atg aat ctc gac cct ttc 4194Pro Ile Leu Phe Ser Gly Ser Leu Arg Met Asn Leu Asp Pro Phe 1385 13901395 aac aac tac tca gat gag gag att tgg aag gcc ttg gag ctg gct 4239Asn Asn Tyr Ser Asp Glu Glu Ile Trp Lys Ala Leu Glu Leu Ala 1400 14051410 cac ctc aag tct ttt gtg gcc agc ctg caa ctt ggg tta tcc cac 4284His Leu Lys Ser Phe Val Ala Ser Leu Gln Leu Gly Leu Ser His 1415 14201425 gaa gtt aca gag gct ggt ggc aac ctg agc ata ggc cag agg cag 4329Glu Val Thr Glu Ala Gly Gly Asn Leu Ser Ile Gly Gln Arg Gln 1430 14351440 ctg ctg tgc ctg ggc agg gct ctg ctt cgg aaa tcc aag atc ctg 4374Leu Leu Cys Leu Gly Arg Ala Leu Leu Arg Lys Ser Lys Ile Leu 1445 14501455 gtc ctg gat gag gcc act gct gcg gtg gat cta gag aca gac aac 4419Val Leu Asp Glu Ala Thr Ala Ala Val Asp Leu Glu Thr Asp Asn 1460 14651470 ctc att cag acg acc atc caa aac gag ttc gcc cac tgc aca gtg 4464Leu Ile Gln Thr Thr Ile Gln Asn Glu Phe Ala His Cys Thr Val 1475 14801485 atc acc atc gcc cac agg ctg cat acc atc atg gac agt gac aag 4509Ile Thr Ile Ala His Arg Leu His Thr Ile Met Asp Ser Asp Lys 1490 14951500 gta atg gtc cta gac aac ggg aag att ata gag tac ggc agc cct 4554Val Met Val Leu Asp Asn Gly Lys Ile Ile Glu Tyr Gly Ser Pro 1505 15101515 gaa gaa ctg cta caa atc cct gga ccc ttt tac ttt atg gct aag 4599Glu Glu Leu Leu Gln Ile Pro Gly Pro Phe Tyr Phe Met Ala Lys 1520 15251530 gaa gct ggc att gag aat gtg aac agc aca aaa gcc gcaccggtcg 4645 GluAla Gly Ile Glu Asn Val Asn Ser Thr Lys Ala 1535 1540 1545 14 1545 PRTHomo sapiens 14 Met Leu Glu Lys Phe Cys Asn Ser Thr Phe Trp Asn Ser SerPhe Leu 1 5 10 15 Asp Ser Pro Glu Ala Asp Leu Pro Leu Cys Phe Glu GlnThr Val Leu 20 25 30 Val Trp Ile Pro Leu Gly Phe Leu Trp Leu Leu Ala ProTrp Gln Leu 35 40 45 Leu His Val Tyr Lys Ser Arg Thr Lys Arg Ser Ser ThrThr Lys Leu 50 55 60 Tyr Leu Ala Lys Gln Val Phe Val Gly Phe Leu Leu IleLeu Ala Ala 65 70 75 80 Ile Glu Leu Ala Leu Val Leu Thr Glu Asp Ser GlyGln Ala Thr Val 85 90 95 Pro Ala Val Arg Tyr Thr Asn Pro Ser Leu Tyr LeuGly Thr Trp Leu 100 105 110 Leu Val Leu Leu Ile Gln Tyr Ser Arg Gln TrpCys Val Gln Lys Asn 115 120 125 Ser Trp Phe Leu Ser Leu Phe Trp Ile LeuSer Ile Leu Cys Gly Thr 130 135 140 Phe Gln Phe Gln Thr Leu Ile Arg ThrLeu Leu Gln Gly Asp Asn Ser 145 150 155 160 Asn Leu Ala Tyr Ser Cys LeuPhe Phe Ile Ser Tyr Gly Phe Gln Ile 165 170 175 Leu Ile Leu Ile Phe SerAla Phe Ser Glu Asn Asn Glu Ser Ser Asn 180 185 190 Asn Pro Ser Ser IleAla Ser Phe Leu Ser Ser Ile Thr Tyr Ser Trp 195 200 205 Tyr Asp Ser IleIle Leu Lys Gly Tyr Lys Arg Pro Leu Thr Leu Glu 210 215 220 Asp Val TrpGlu Val Asp Glu Glu Met Lys Thr Lys Thr Leu Val Ser 225 230 235 240 LysPhe Glu Thr His Met Lys Arg Glu Leu Gln Lys Ala Arg Arg Ala 245 250 255Leu Gln Arg Arg Gln Glu Lys Ser Ser Gln Gln Asn Ser Gly Ala Arg 260 265270 Leu Pro Gly Leu Asn Lys Asn Gln Ser Gln Ser Gln Asp Ala Leu Val 275280 285 Leu Glu Asp Val Glu Lys Lys Lys Lys Lys Ser Gly Thr Lys Lys Asp290 295 300 Val Pro Lys Ser Trp Leu Met Lys Ala Leu Phe Lys Thr Phe TyrMet 305 310 315 320 Val Leu Leu Lys Ser Phe Leu Leu Lys Leu Val Asn AspIle Phe Thr 325 330 335 Phe Val Ser Pro Gln Leu Leu Lys Leu Leu Ile SerPhe Ala Ser Asp 340 345 350 Arg Asp Thr Tyr Leu Trp Ile Gly Tyr Leu CysAla Ile Leu Leu Phe 355 360 365 Thr Ala Ala Leu Ile Gln Ser Phe Cys LeuGln Cys Tyr Phe Gln Leu 370 375 380 Cys Phe Lys Leu Gly Val Lys Val ArgThr Ala Ile Met Ala Ser Val 385 390 395 400 Tyr Lys Lys Ala Leu Thr LeuSer Asn Leu Ala Arg Lys Glu Tyr Thr 405 410 415 Val Gly Glu Thr Val AsnLeu Met Ser Val Asp Ala Gln Lys Leu Met 420 425 430 Asp Val Thr Asn PheMet His Met Leu Trp Ser Ser Val Leu Gln Ile 435 440 445 Val Leu Ser IlePhe Phe Leu Trp Arg Glu Leu Gly Pro Ser Val Leu 450 455 460 Ala Gly ValGly Val Met Val Leu Val Ile Pro Ile Asn Ala Ile Leu 465 470 475 480 SerThr Lys Ser Lys Thr Ile Gln Val Lys Asn Met Lys Asn Lys Asp 485 490 495Lys Arg Leu Lys Ile Met Asn Glu Ile Leu Ser Gly Ile Lys Ile Leu 500 505510 Lys Tyr Phe Ala Trp Glu Pro Ser Phe Arg Asp Gln Val Gln Asn Leu 515520 525 Arg Lys Lys Glu Leu Lys Asn Leu Leu Ala Phe Ser Gln Leu Gln Cys530 535 540 Val Val Ile Phe Val Phe Gln Leu Thr Pro Val Leu Val Ser ValVal 545 550 555 560 Thr Phe Ser Val Tyr Val Leu Val Asp Ser Asn Asn IleLeu Asp Ala 565 570 575 Gln Lys Ala Phe Thr Ser Ile Thr Leu Phe Asn IleLeu Arg Phe Pro 580 585 590 Leu Ser Met Leu Pro Met Met Ile Ser Ser MetLeu Gln Ala Ser Val 595 600 605 Ser Thr Glu Arg Leu Glu Lys Tyr Leu GlyGly Asp Asp Leu Asp Thr 610 615 620 Ser Ala Ile Arg His Asp Cys Asn PheAsp Lys Ala Met Gln Phe Ser 625 630 635 640 Glu Ala Ser Phe Thr Trp GluHis Asp Ser Glu Ala Thr Val Arg Asp 645 650 655 Val Asn Leu Asp Ile MetAla Gly Gln Leu Val Ala Val Ile Gly Pro 660 665 670 Val Gly Ser Gly LysSer Ser Leu Ile Ser Ala Met Leu Gly Glu Met 675 680 685 Glu Asn Val HisGly His Ile Thr Ile Lys Gly Thr Thr Ala Tyr Val 690 695 700 Pro Gln GlnSer Trp Ile Gln Asn Gly Thr Ile Lys Asp Asn Ile Leu 705 710 715 720 PheGly Thr Glu Phe Asn Glu Lys Arg Tyr Gln Gln Val Leu Glu Ala 725 730 735Cys Ala Leu Leu Pro Asp Leu Glu Met Leu Pro Gly Gly Asp Leu Ala 740 745750 Glu Ile Gly Glu Lys Gly Ile Asn Leu Ser Gly Gly Gln Lys Gln Arg 755760 765 Ile Ser Leu Ala Arg Ala Thr Tyr Gln Asn Leu Asp Ile Tyr Leu Leu770 775 780 Asp Asp Pro Leu Ser Ala Val Asp Ala His Val Gly Lys His IlePhe 785 790 795 800 Asn Lys Val Leu Gly Pro Asn Gly Leu Leu Lys Gly LysThr Arg Leu 805 810 815 Leu Val Thr His Ser Met His Phe Leu Pro Gln ValAsp Glu Ile Val 820 825 830 Val Leu Gly Asn Gly Thr Ile Val Glu Lys GlySer Tyr Ser Ala Leu 835 840 845 Leu Ala Lys Lys Gly Glu Phe Ala Lys AsnLeu Lys Thr Phe Leu Arg 850 855 860 His Thr Gly Pro Glu Glu Glu Ala ThrVal His Asp Gly Ser Glu Glu 865 870 875 880 Glu Asp Asp Asp Tyr Gly LeuIle Ser Ser Val Glu Glu Ile Pro Glu 885 890 895 Asp Ala Ala Ser Ile ThrMet Arg Arg Glu Asn Ser Phe Arg Arg Thr 900 905 910 Leu Ser Arg Ser SerArg Ser Asn Gly Arg His Leu Lys Ser Leu Arg 915 920 925 Asn Ser Leu LysThr Arg Asn Val Asn Ser Leu Lys Glu Asp Glu Glu 930 935 940 Leu Val LysGly Gln Lys Leu Ile Lys Lys Glu Phe Ile Glu Thr Gly 945 950 955 960 LysVal Lys Phe Ser Ile Tyr Leu Glu Tyr Leu Gln Ala Ile Gly Leu 965 970 975Phe Ser Ile Phe Phe Ile Ile Leu Ala Phe Val Met Asn Ser Val Ala 980 985990 Phe Ile Gly Ser Asn Leu Trp Leu Ser Ala Trp Thr Ser Asp Ser Lys 9951000 1005 Ile Phe Asn Ser Thr Asp Tyr Pro Ala Ser Gln Arg Asp Met Arg1010 1015 1020 Val Gly Val Tyr Gly Ala Leu Gly Leu Ala Gln Gly Ile PheVal 1025 1030 1035 Phe Ile Ala His Phe Trp Ser Ala Phe Gly Phe Val HisAla Ser 1040 1045 1050 Asn Ile Leu His Lys Gln Leu Leu Asn Asn Ile LeuArg Ala Pro 1055 1060 1065 Met Arg Phe Phe Asp Thr Thr Pro Thr Gly ArgIle Val Asn Arg 1070 1075 1080 Phe Ala Gly Asp Ile Ser Thr Val Asp AspThr Leu Pro Gln Ser 1085 1090 1095 Leu Arg Ser Trp Ile Thr Cys Phe LeuGly Ile Ile Ser Thr Leu 1100 1105 1110 Val Met Ile Cys Met Ala Thr ProVal Phe Thr Ile Ile Val Ile 1115 1120 1125 Pro Leu Gly Ile Ile Tyr ValSer Val Gln Met Phe Tyr Val Ser 1130 1135 1140 Thr Ser Arg Gln Leu ArgArg Leu Asp Ser Val Thr Arg Ser Pro 1145 1150 1155 Ile Tyr Ser His PheSer Glu Thr Val Ser Gly Leu Pro Val Ile 1160 1165 1170 Arg Ala Phe GluHis Gln Gln Arg Phe Leu Lys His Asn Glu Glu 1175 1180 1185 Arg Ile AspThr Asn Gln Lys Cys Val Phe Ser Trp Ile Thr Ser 1190 1195 1200 Asn ArgTrp Leu Ala Ile Arg Leu Glu Leu Val Gly Asn Leu Thr 1205 1210 1215 ValPhe Phe Ser Ala Leu Met Met Val Ile Tyr Arg Asp Thr Leu 1220 1225 1230Ser Gly Asp Thr Val Gly Phe Val Leu Ser Asn Ala Leu Asn Ile 1235 12401245 Thr Gln Thr Leu Asn Trp Leu Val Arg Met Thr Ser Glu Ile Glu 12501255 1260 Thr Asn Ile Val Ala Val Glu Arg Ile Thr Glu Tyr Thr Lys Val1265 1270 1275 Glu Asn Glu Ala Pro Trp Val Thr Asp Lys Arg Pro Pro ProAsp 1280 1285 1290 Trp Pro Ser Lys Gly Lys Ile Gln Phe Asn Asn Tyr GlnVal Arg 1295 1300 1305 Tyr Arg Pro Glu Leu Asp Leu Val Leu Arg Gly IleThr Cys Asp 1310 1315 1320 Ile Gly Ser Met Glu Lys Ile Gly Val Val GlyArg Thr Gly Ala 1325 1330 1335 Gly Lys Ser Ser Leu Thr Asn Cys Leu PheArg Ile Leu Glu Ala 1340 1345 1350 Ala Gly Gly Gln Ile Ile Ile Asp GlyVal Asp Ile Ala Ser Ile 1355 1360 1365 Gly Leu His Asp Leu Arg Glu LysLeu Thr Ile Ile Pro Gln Asp 1370 1375 1380 Pro Ile Leu Phe Ser Gly SerLeu Arg Met Asn Leu Asp Pro Phe 1385 1390 1395 Asn Asn Tyr Ser Asp GluGlu Ile Trp Lys Ala Leu Glu Leu Ala 1400 1405 1410 His Leu Lys Ser PheVal Ala Ser Leu Gln Leu Gly Leu Ser His 1415 1420 1425 Glu Val Thr GluAla Gly Gly Asn Leu Ser Ile Gly Gln Arg Gln 1430 1435 1440 Leu Leu CysLeu Gly Arg Ala Leu Leu Arg Lys Ser Lys Ile Leu 1445 1450 1455 Val LeuAsp Glu Ala Thr Ala Ala Val Asp Leu Glu Thr Asp Asn 1460 1465 1470 LeuIle Gln Thr Thr Ile Gln Asn Glu Phe Ala His Cys Thr Val 1475 1480 1485Ile Thr Ile Ala His Arg Leu His Thr Ile Met Asp Ser Asp Lys 1490 14951500 Val Met Val Leu Asp Asn Gly Lys Ile Ile Glu Tyr Gly Ser Pro 15051510 1515 Glu Glu Leu Leu Gln Ile Pro Gly Pro Phe Tyr Phe Met Ala Lys1520 1525 1530 Glu Ala Gly Ile Glu Asn Val Asn Ser Thr Lys Ala 1535 15401545 15 4645 DNA Homo sapiens CDS (1)..(4635) 15 atg ctg gag aag ttc tgcaac tct act ttt tgg aat tcc tca ttc ctg 48 Met Leu Glu Lys Phe Cys AsnSer Thr Phe Trp Asn Ser Ser Phe Leu 1 5 10 15 gac agt ccg gag gca gacctg cca ctt tgt ttt gag caa act gtt ctg 96 Asp Ser Pro Glu Ala Asp LeuPro Leu Cys Phe Glu Gln Thr Val Leu 20 25 30 gtg tgg att ccc ttg ggc ttccta tgg ctc ctg gcc ccc tgg cag ctt 144 Val Trp Ile Pro Leu Gly Phe LeuTrp Leu Leu Ala Pro Trp Gln Leu 35 40 45 ctc cac gtg tat aaa tcc agg accaag aga tcc tct acc acc aaa ctc 192 Leu His Val Tyr Lys Ser Arg Thr LysArg Ser Ser Thr Thr Lys Leu 50 55 60 tat ctt gct aag cag gta ttc gtt ggtttt ctt ctt att cta gca gcc 240 Tyr Leu Ala Lys Gln Val Phe Val Gly PheLeu Leu Ile Leu Ala Ala 65 70 75 80 ata gag ctg gcc ctt gta ctc aca gaagac tct gga caa gcc aca gtc 288 Ile Glu Leu Ala Leu Val Leu Thr Glu AspSer Gly Gln Ala Thr Val 85 90 95 cct gct gtt cga tat acc aat cca agc ctctac cta ggc aca tgg ctc 336 Pro Ala Val Arg Tyr Thr Asn Pro Ser Leu TyrLeu Gly Thr Trp Leu 100 105 110 ctg gtt ttg ctg atc caa tac agc aga caatgg tgt gta cag aaa aac 384 Leu Val Leu Leu Ile Gln Tyr Ser Arg Gln TrpCys Val Gln Lys Asn 115 120 125 tcc tgg ttc ctg tcc cta ttc tgg att ctctcg ata ctc tgt ggc act 432 Ser Trp Phe Leu Ser Leu Phe Trp Ile Leu SerIle Leu Cys Gly Thr 130 135 140 ttc caa ttt cag act ctg atc cgg aca ctctta cag ggt gac aat tct 480 Phe Gln Phe Gln Thr Leu Ile Arg Thr Leu LeuGln Gly Asp Asn Ser 145 150 155 160 aat cta gcc tac tcc tgc ctg ttc ttcatc tcc tac gga ttc cag atc 528 Asn Leu Ala Tyr Ser Cys Leu Phe Phe IleSer Tyr Gly Phe Gln Ile 165 170 175 ctg atc ctg atc ttt tca gca ttt tcagaa aat aat gag tca tca aat 576 Leu Ile Leu Ile Phe Ser Ala Phe Ser GluAsn Asn Glu Ser Ser Asn 180 185 190 aat cca tca tcc ata gct tca ttc ctgagt agc att acc tac agc tgg 624 Asn Pro Ser Ser Ile Ala Ser Phe Leu SerSer Ile Thr Tyr Ser Trp 195 200 205 tat gac agc atc att ctg aaa ggc tacaag cgt cct ctg aca ctc gag 672 Tyr Asp Ser Ile Ile Leu Lys Gly Tyr LysArg Pro Leu Thr Leu Glu 210 215 220 gat gtc tgg gaa gtt gat gaa gag atgaaa acc aag aca tta gtg agc 720 Asp Val Trp Glu Val Asp Glu Glu Met LysThr Lys Thr Leu Val Ser 225 230 235 240 aag ttt gaa acg cac atg aag agagag ctg cag aaa gcc agg cgg gca 768 Lys Phe Glu Thr His Met Lys Arg GluLeu Gln Lys Ala Arg Arg Ala 245 250 255 ctc cag aga cgg cag gag aag agctcc cag cag aac tct gga gcc agg 816 Leu Gln Arg Arg Gln Glu Lys Ser SerGln Gln Asn Ser Gly Ala Arg 260 265 270 ctg cct ggc ttg aac aag aat cagagt caa agc caa gat gcc ctt gtc 864 Leu Pro Gly Leu Asn Lys Asn Gln SerGln Ser Gln Asp Ala Leu Val 275 280 285 ctg gaa gat gtt gaa aag aaa aaaaag aag tct ggg acc aaa aaa gat 912 Leu Glu Asp Val Glu Lys Lys Lys LysLys Ser Gly Thr Lys Lys Asp 290 295 300 gtt cca aaa tcc tgg ttg atg aaggct ctg ttc aaa act ttc tac atg 960 Val Pro Lys Ser Trp Leu Met Lys AlaLeu Phe Lys Thr Phe Tyr Met 305 310 315 320 gtg ctc ctg aaa tca ttc ctactg aag cta gtg aat gac atc ttc acg 1008 Val Leu Leu Lys Ser Phe Leu LeuLys Leu Val Asn Asp Ile Phe Thr 325 330 335 ttt gtg agt cct cag ctg ctgaaa ttg ctg atc tcc ttt gca agt gac 1056 Phe Val Ser Pro Gln Leu Leu LysLeu Leu Ile Ser Phe Ala Ser Asp 340 345 350 cgt gac aca tat ttg tgg attgga tat ctc tgt gca atc ctc tta ttc 1104 Arg Asp Thr Tyr Leu Trp Ile GlyTyr Leu Cys Ala Ile Leu Leu Phe 355 360 365 act gcg gct ctc att cag tctttc tgc ctt cag tgt tat ttc caa ctg 1152 Thr Ala Ala Leu Ile Gln Ser PheCys Leu Gln Cys Tyr Phe Gln Leu 370 375 380 tgc ttc aag ctg ggt gta aaagta cgg aca gct atc atg gct tct gta 1200 Cys Phe Lys Leu Gly Val Lys ValArg Thr Ala Ile Met Ala Ser Val 385 390 395 400 tat aag aag gca ttg acccta tcc aac ttg gcc agg aag gag tac acc 1248 Tyr Lys Lys Ala Leu Thr LeuSer Asn Leu Ala Arg Lys Glu Tyr Thr 405 410 415 gtt gga gaa aca gtg aacctg atg tct gtg gat gcc cag aag ctc atg 1296 Val Gly Glu Thr Val Asn LeuMet Ser Val Asp Ala Gln Lys Leu Met 420 425 430 gat gtg acc aac ttc atgcac atg ctg tgg tca agt gtt cta cag att 1344 Asp Val Thr Asn Phe Met HisMet Leu Trp Ser Ser Val Leu Gln Ile 435 440 445 gtc tta tct atc ttc ttccta tgg aga gag ttg gga ccc tca gtc tta 1392 Val Leu Ser Ile Phe Phe LeuTrp Arg Glu Leu Gly Pro Ser Val Leu 450 455 460 gca ggt gtt ggg gtg atggtg ctt gta atc cca att aat gcg ata ctg 1440 Ala Gly Val Gly Val Met ValLeu Val Ile Pro Ile Asn Ala Ile Leu 465 470 475 480 tcc acc aag agt aagacc att cag gtc aaa aat atg aag aat aaa gac 1488 Ser Thr Lys Ser Lys ThrIle Gln Val Lys Asn Met Lys Asn Lys Asp 485 490 495 aaa cgt tta aag atcatg aat gag att ctt agt gga atc aag atc ctg 1536 Lys Arg Leu Lys Ile MetAsn Glu Ile Leu Ser Gly Ile Lys Ile Leu 500 505 510 aaa tat ttt gcc tgggaa cct tca ttc aga gac caa gta caa aac ctc 1584 Lys Tyr Phe Ala Trp GluPro Ser Phe Arg Asp Gln Val Gln Asn Leu 515 520 525 cgg aag aaa gag ctcaag aac ctg ctg gcc ttt agt caa cta cag tgt 1632 Arg Lys Lys Glu Leu LysAsn Leu Leu Ala Phe Ser Gln Leu Gln Cys 530 535 540 gta gta ata ttc gtcttc cag tta act cca gtc ctg gta tct gtg gtc 1680 Val Val Ile Phe Val PheGln Leu Thr Pro Val Leu Val Ser Val Val 545 550 555 560 aca ttt tct gtttat gtc ctg gtg gat agc aac aat att ttg gat gca 1728 Thr Phe Ser Val TyrVal Leu Val Asp Ser Asn Asn Ile Leu Asp Ala 565 570 575 caa aag gcc ttcacc tcc att acc ctc ttc aat atc ctg cgc ttt ccc 1776 Gln Lys Ala Phe ThrSer Ile Thr Leu Phe Asn Ile Leu Arg Phe Pro 580 585 590 ctg agc atg cttccc atg atg atc tcc tcc atg ctc cag gcc agt gtt 1824 Leu Ser Met Leu ProMet Met Ile Ser Ser Met Leu Gln Ala Ser Val 595 600 605 tcc aca gag cggcta gag aag tac ttg gga ggg gat gac ttg gac aca 1872 Ser Thr Glu Arg LeuGlu Lys Tyr Leu Gly Gly Asp Asp Leu Asp Thr 610 615 620 tct gcc att cgacat gac tgc aat ttt gac aaa gcc atg cag ttt tct 1920 Ser Ala Ile Arg HisAsp Cys Asn Phe Asp Lys Ala Met Gln Phe Ser 625 630 635 640 gag gcc tccttt acc tgg gaa cat gat tcg gaa gcc aca gtc cga gat 1968 Glu Ala Ser PheThr Trp Glu His Asp Ser Glu Ala Thr Val Arg Asp 645 650 655 gtg aac ctggac att atg gca ggc caa ctt gtg gct gtg ata ggc cct 2016 Val Asn Leu AspIle Met Ala Gly Gln Leu Val Ala Val Ile Gly Pro 660 665 670 gtc ggc tctggg aaa tcc tcc ttg ata tca gcc atg ctg gga gaa atg 2064 Val Gly Ser GlyLys Ser Ser Leu Ile Ser Ala Met Leu Gly Glu Met 675 680 685 gaa aat gtccac ggg cac atc acc atc aag ggc acc act gcc tat gtc 2112 Glu Asn Val HisGly His Ile Thr Ile Lys Gly Thr Thr Ala Tyr Val 690 695 700 cca cag cagtcc tgg att cag aat ggc acc ata aag gac aac atc ctt 2160 Pro Gln Gln SerTrp Ile Gln Asn Gly Thr Ile Lys Asp Asn Ile Leu 705 710 715 720 ttt ggaaca gag ttt aat gaa aag agg tac cag caa gta ctg gag gcc 2208 Phe Gly ThrGlu Phe Asn Glu Lys Arg Tyr Gln Gln Val Leu Glu Ala 725 730 735 tgt gctctc ctc cca gac ttg gaa atg ctg cct gga gga gat ttg gct 2256 Cys Ala LeuLeu Pro Asp Leu Glu Met Leu Pro Gly Gly Asp Leu Ala 740 745 750 gag attgga gag aag ggt ata aat ctt agt ggg ggt cag aag cag cgg 2304 Glu Ile GlyGlu Lys Gly Ile Asn Leu Ser Gly Gly Gln Lys Gln Arg 755 760 765 atc agcctg gcc aga gct acc tac caa aat tta gac atc tat ctt cta 2352 Ile Ser LeuAla Arg Ala Thr Tyr Gln Asn Leu Asp Ile Tyr Leu Leu 770 775 780 gat gacccc ctg tct gca gtg gat gct cat gta gga aaa cat att ttt 2400 Asp Asp ProLeu Ser Ala Val Asp Ala His Val Gly Lys His Ile Phe 785 790 795 800 aataag gtc ttg ggc ccc aat ggc ctg ttg aaa ggc aag act cga ctc 2448 Asn LysVal Leu Gly Pro Asn Gly Leu Leu Lys Gly Lys Thr Arg Leu 805 810 815 ttggtt aca cat agc atg cac ttt ctt cct caa gtg gat gag att gta 2496 Leu ValThr His Ser Met His Phe Leu Pro Gln Val Asp Glu Ile Val 820 825 830 gttctg ggg aat gga aca att gta gag aaa gga tcc tac agt gct ctc 2544 Val LeuGly Asn Gly Thr Ile Val Glu Lys Gly Ser Tyr Ser Ala Leu 835 840 845 ctggcc aaa aaa gga gag ttt gct aag aat ctg aag aca ttt cta aga 2592 Leu AlaLys Lys Gly Glu Phe Ala Lys Asn Leu Lys Thr Phe Leu Arg 850 855 860 cataca ggc cct gaa gag gaa gcc aca gtc cat gat ggc agt gaa gaa 2640 His ThrGly Pro Glu Glu Glu Ala Thr Val His Asp Gly Ser Glu Glu 865 870 875 880gaa gac gat gac tat ggg ctg ata tcc agt gtg gaa gag atc ccc gaa 2688 GluAsp Asp Asp Tyr Gly Leu Ile Ser Ser Val Glu Glu Ile Pro Glu 885 890 895gat gca gcc tcc ata acc atg aga aga gag aac agc ttt cgt cga aca 2736 AspAla Ala Ser Ile Thr Met Arg Arg Glu Asn Ser Phe Arg Arg Thr 900 905 910ctt agc cgc agt tct agg tcc aat ggc agg cat ctg aag tcc ctg aga 2784 LeuSer Arg Ser Ser Arg Ser Asn Gly Arg His Leu Lys Ser Leu Arg 915 920 925aac tcc ttg aaa act cgg aat gtg aat agc ctg aag gaa gac gaa gaa 2832 AsnSer Leu Lys Thr Arg Asn Val Asn Ser Leu Lys Glu Asp Glu Glu 930 935 940cta gtg aaa gga caa aaa cta att aag aag gaa ttc ata gaa act gga 2880 LeuVal Lys Gly Gln Lys Leu Ile Lys Lys Glu Phe Ile Glu Thr Gly 945 950 955960 aag gtg aag ttc tcc atc tac ctg gag tac cta caa gca ata gga ttg 2928Lys Val Lys Phe Ser Ile Tyr Leu Glu Tyr Leu Gln Ala Ile Gly Leu 965 970975 ttt tcg ata ttc ttc atc atc ctt gcg ttt gtg atg aat tct gtg gct 2976Phe Ser Ile Phe Phe Ile Ile Leu Ala Phe Val Met Asn Ser Val Ala 980 985990 ttt att gga tcc aac ctc tgg ctc agt gct tgg acc agt gac tct aaa 3024Phe Ile Gly Ser Asn Leu Trp Leu Ser Ala Trp Thr Ser Asp Ser Lys 995 10001005 atc ttc aat agc acc gac tat cca gca tct cag agg gac atg aga 3069Ile Phe Asn Ser Thr Asp Tyr Pro Ala Ser Gln Arg Asp Met Arg 1010 10151020 gtt gga gtc tac gga gct ctg gga tta gcc caa ggt ata ttt gtg 3114Val Gly Val Tyr Gly Ala Leu Gly Leu Ala Gln Gly Ile Phe Val 1025 10301035 ttc ata gca cat ttc tgg agt gcc ttt ggt ttc gtc cat gca tca 3159Phe Ile Ala His Phe Trp Ser Ala Phe Gly Phe Val His Ala Ser 1040 10451050 aat atc ttg cac aag caa ctg ctg aac aat atc ctt cga gca cct 3204Asn Ile Leu His Lys Gln Leu Leu Asn Asn Ile Leu Arg Ala Pro 1055 10601065 atg aga ttt ttt gac aca aca ccc aca ggc cgg att gtg aac agg 3249Met Arg Phe Phe Asp Thr Thr Pro Thr Gly Arg Ile Val Asn Arg 1070 10751080 ttt gcc ggc gat att tcc aca gtg gat gac acc ctg cct cag tcc 3294Phe Ala Gly Asp Ile Ser Thr Val Asp Asp Thr Leu Pro Gln Ser 1085 10901095 ttg cgc agc tgg att aca tgc ttc ctg ggg ata atc agc acc ctt 3339Leu Arg Ser Trp Ile Thr Cys Phe Leu Gly Ile Ile Ser Thr Leu 1100 11051110 gtc atg atc tgc atg gcc act cct gtc ttc acc atc atc gtc att 3384Val Met Ile Cys Met Ala Thr Pro Val Phe Thr Ile Ile Val Ile 1115 11201125 cct ctt ggc att att tat gta tct gtt cag atg ttt tat gtg tct 3429Pro Leu Gly Ile Ile Tyr Val Ser Val Gln Met Phe Tyr Val Ser 1130 11351140 acc tcc cgc cag ctg agg cgt ctg gac tct gtc acc agg tcc cca 3474Thr Ser Arg Gln Leu Arg Arg Leu Asp Ser Val Thr Arg Ser Pro 1145 11501155 atc tac tct cac ttc agc gag acc gta tca ggt ttg cca gtt atc 3519Ile Tyr Ser His Phe Ser Glu Thr Val Ser Gly Leu Pro Val Ile 1160 11651170 cgt gcc ttt gag cac cag cag cga ttt ctg aaa cac aat gag gag 3564Arg Ala Phe Glu His Gln Gln Arg Phe Leu Lys His Asn Glu Glu 1175 11801185 agg att gac acc aac cag aaa tgt gtc ttt tcc tgg atc acc tcc 3609Arg Ile Asp Thr Asn Gln Lys Cys Val Phe Ser Trp Ile Thr Ser 1190 11951200 aac agg tgg ctt gca att cgc ctg gag ctg gtt ggg aac ctg act 3654Asn Arg Trp Leu Ala Ile Arg Leu Glu Leu Val Gly Asn Leu Thr 1205 12101215 gtc ttc ttt tca gcc ttg atg atg gtt att tat aga gat acc cta 3699Val Phe Phe Ser Ala Leu Met Met Val Ile Tyr Arg Asp Thr Leu 1220 12251230 agt ggg gac act gtt ggc ttt gtt ctg tcc aat gca ctc aat atc 3744Ser Gly Asp Thr Val Gly Phe Val Leu Ser Asn Ala Leu Asn Ile 1235 12401245 aca caa acc ctg aac tgg ctg gtg agg atg aca tca gaa ata gag 3789Thr Gln Thr Leu Asn Trp Leu Val Arg Met Thr Ser Glu Ile Glu 1250 12551260 acc aac att gtg gct gtt gag cga ata act gag tac aca aaa gtg 3834Thr Asn Ile Val Ala Val Glu Arg Ile Thr Glu Tyr Thr Lys Val 1265 12701275 gaa aat gag gca ccc tgg gtg act gat aag agg cct ccg cca gat 3879Glu Asn Glu Ala Pro Trp Val Thr Asp Lys Arg Pro Pro Pro Asp 1280 12851290 tgg ccc agc aaa ggc aag atc cag ttt aac aac tac caa gtg cgg 3924Trp Pro Ser Lys Gly Lys Ile Gln Phe Asn Asn Tyr Gln Val Arg 1295 13001305 tac cga cct gag ctg gat ctg gtc ctc aga ggg atc act tgt gac 3969Tyr Arg Pro Glu Leu Asp Leu Val Leu Arg Gly Ile Thr Cys Asp 1310 13151320 atc ggt agc atg gag aag att ggt gtg gtg ggc agg aca gga gct 4014Ile Gly Ser Met Glu Lys Ile Gly Val Val Gly Arg Thr Gly Ala 1325 13301335 gga aag tca tcc ctc aca aac tgc ctc ttc aga atc tta gag gct 4059Gly Lys Ser Ser Leu Thr Asn Cys Leu Phe Arg Ile Leu Glu Ala 1340 13451350 gcc ggt ggt cag att atc att gat gga gta gat att gct tcc att 4104Ala Gly Gly Gln Ile Ile Ile Asp Gly Val Asp Ile Ala Ser Ile 1355 13601365 ggg ctc cac gac ctc cga gag aag ctg acc atc atc ccc cag gac 4149Gly Leu His Asp Leu Arg Glu Lys Leu Thr Ile Ile Pro Gln Asp 1370 13751380 ccc atc ctg ttc tct gga agc ctg agg atg aat ctc gac cct ttc 4194Pro Ile Leu Phe Ser Gly Ser Leu Arg Met Asn Leu Asp Pro Phe 1385 13901395 aac aac tac tca gat gag gag att tgg aag gcc ttg gag ctg gct 4239Asn Asn Tyr Ser Asp Glu Glu Ile Trp Lys Ala Leu Glu Leu Ala 1400 14051410 cac ctc aag tct ttt gtg gcc agc ctg caa ctt ggg tta tcc cac 4284His Leu Lys Ser Phe Val Ala Ser Leu Gln Leu Gly Leu Ser His 1415 14201425 gaa gtt aca gag gct ggt ggc aac ctg agc ata ggc cag agg cag 4329Glu Val Thr Glu Ala Gly Gly Asn Leu Ser Ile Gly Gln Arg Gln 1430 14351440 ctg ctg tgc ctg ggc agg gct ctg ctt cgg aaa tcc aag atc ctg 4374Leu Leu Cys Leu Gly Arg Ala Leu Leu Arg Lys Ser Lys Ile Leu 1445 14501455 gtc ctg gat gag gcc act gct gcg gtg gat cta gag aca gac aac 4419Val Leu Asp Glu Ala Thr Ala Ala Val Asp Leu Glu Thr Asp Asn 1460 14651470 ctc att cag acg acc atc caa aac gag ttc gcc cac tgc aca gtg 4464Leu Ile Gln Thr Thr Ile Gln Asn Glu Phe Ala His Cys Thr Val 1475 14801485 atc acc atc gcc cac agg ctg cat acc atc atg gac agt gac aag 4509Ile Thr Ile Ala His Arg Leu His Thr Ile Met Asp Ser Asp Lys 1490 14951500 gta atg gtc cta gac aac ggg aag att ata gag tac ggc agc cct 4554Val Met Val Leu Asp Asn Gly Lys Ile Ile Glu Tyr Gly Ser Pro 1505 15101515 gaa gaa ctg cta caa atc cct gga ccc ttt tac ttt atg gct aag 4599Glu Glu Leu Leu Gln Ile Pro Gly Pro Phe Tyr Phe Met Ala Lys 1520 15251530 gaa gct ggc att gag aat gtg aac agc gca gca gcc gcaccggtcg 4645 GluAla Gly Ile Glu Asn Val Asn Ser Ala Ala Ala 1535 1540 1545 16 1545 PRTHomo sapiens 16 Met Leu Glu Lys Phe Cys Asn Ser Thr Phe Trp Asn Ser SerPhe Leu 1 5 10 15 Asp Ser Pro Glu Ala Asp Leu Pro Leu Cys Phe Glu GlnThr Val Leu 20 25 30 Val Trp Ile Pro Leu Gly Phe Leu Trp Leu Leu Ala ProTrp Gln Leu 35 40 45 Leu His Val Tyr Lys Ser Arg Thr Lys Arg Ser Ser ThrThr Lys Leu 50 55 60 Tyr Leu Ala Lys Gln Val Phe Val Gly Phe Leu Leu IleLeu Ala Ala 65 70 75 80 Ile Glu Leu Ala Leu Val Leu Thr Glu Asp Ser GlyGln Ala Thr Val 85 90 95 Pro Ala Val Arg Tyr Thr Asn Pro Ser Leu Tyr LeuGly Thr Trp Leu 100 105 110 Leu Val Leu Leu Ile Gln Tyr Ser Arg Gln TrpCys Val Gln Lys Asn 115 120 125 Ser Trp Phe Leu Ser Leu Phe Trp Ile LeuSer Ile Leu Cys Gly Thr 130 135 140 Phe Gln Phe Gln Thr Leu Ile Arg ThrLeu Leu Gln Gly Asp Asn Ser 145 150 155 160 Asn Leu Ala Tyr Ser Cys LeuPhe Phe Ile Ser Tyr Gly Phe Gln Ile 165 170 175 Leu Ile Leu Ile Phe SerAla Phe Ser Glu Asn Asn Glu Ser Ser Asn 180 185 190 Asn Pro Ser Ser IleAla Ser Phe Leu Ser Ser Ile Thr Tyr Ser Trp 195 200 205 Tyr Asp Ser IleIle Leu Lys Gly Tyr Lys Arg Pro Leu Thr Leu Glu 210 215 220 Asp Val TrpGlu Val Asp Glu Glu Met Lys Thr Lys Thr Leu Val Ser 225 230 235 240 LysPhe Glu Thr His Met Lys Arg Glu Leu Gln Lys Ala Arg Arg Ala 245 250 255Leu Gln Arg Arg Gln Glu Lys Ser Ser Gln Gln Asn Ser Gly Ala Arg 260 265270 Leu Pro Gly Leu Asn Lys Asn Gln Ser Gln Ser Gln Asp Ala Leu Val 275280 285 Leu Glu Asp Val Glu Lys Lys Lys Lys Lys Ser Gly Thr Lys Lys Asp290 295 300 Val Pro Lys Ser Trp Leu Met Lys Ala Leu Phe Lys Thr Phe TyrMet 305 310 315 320 Val Leu Leu Lys Ser Phe Leu Leu Lys Leu Val Asn AspIle Phe Thr 325 330 335 Phe Val Ser Pro Gln Leu Leu Lys Leu Leu Ile SerPhe Ala Ser Asp 340 345 350 Arg Asp Thr Tyr Leu Trp Ile Gly Tyr Leu CysAla Ile Leu Leu Phe 355 360 365 Thr Ala Ala Leu Ile Gln Ser Phe Cys LeuGln Cys Tyr Phe Gln Leu 370 375 380 Cys Phe Lys Leu Gly Val Lys Val ArgThr Ala Ile Met Ala Ser Val 385 390 395 400 Tyr Lys Lys Ala Leu Thr LeuSer Asn Leu Ala Arg Lys Glu Tyr Thr 405 410 415 Val Gly Glu Thr Val AsnLeu Met Ser Val Asp Ala Gln Lys Leu Met 420 425 430 Asp Val Thr Asn PheMet His Met Leu Trp Ser Ser Val Leu Gln Ile 435 440 445 Val Leu Ser IlePhe Phe Leu Trp Arg Glu Leu Gly Pro Ser Val Leu 450 455 460 Ala Gly ValGly Val Met Val Leu Val Ile Pro Ile Asn Ala Ile Leu 465 470 475 480 SerThr Lys Ser Lys Thr Ile Gln Val Lys Asn Met Lys Asn Lys Asp 485 490 495Lys Arg Leu Lys Ile Met Asn Glu Ile Leu Ser Gly Ile Lys Ile Leu 500 505510 Lys Tyr Phe Ala Trp Glu Pro Ser Phe Arg Asp Gln Val Gln Asn Leu 515520 525 Arg Lys Lys Glu Leu Lys Asn Leu Leu Ala Phe Ser Gln Leu Gln Cys530 535 540 Val Val Ile Phe Val Phe Gln Leu Thr Pro Val Leu Val Ser ValVal 545 550 555 560 Thr Phe Ser Val Tyr Val Leu Val Asp Ser Asn Asn IleLeu Asp Ala 565 570 575 Gln Lys Ala Phe Thr Ser Ile Thr Leu Phe Asn IleLeu Arg Phe Pro 580 585 590 Leu Ser Met Leu Pro Met Met Ile Ser Ser MetLeu Gln Ala Ser Val 595 600 605 Ser Thr Glu Arg Leu Glu Lys Tyr Leu GlyGly Asp Asp Leu Asp Thr 610 615 620 Ser Ala Ile Arg His Asp Cys Asn PheAsp Lys Ala Met Gln Phe Ser 625 630 635 640 Glu Ala Ser Phe Thr Trp GluHis Asp Ser Glu Ala Thr Val Arg Asp 645 650 655 Val Asn Leu Asp Ile MetAla Gly Gln Leu Val Ala Val Ile Gly Pro 660 665 670 Val Gly Ser Gly LysSer Ser Leu Ile Ser Ala Met Leu Gly Glu Met 675 680 685 Glu Asn Val HisGly His Ile Thr Ile Lys Gly Thr Thr Ala Tyr Val 690 695 700 Pro Gln GlnSer Trp Ile Gln Asn Gly Thr Ile Lys Asp Asn Ile Leu 705 710 715 720 PheGly Thr Glu Phe Asn Glu Lys Arg Tyr Gln Gln Val Leu Glu Ala 725 730 735Cys Ala Leu Leu Pro Asp Leu Glu Met Leu Pro Gly Gly Asp Leu Ala 740 745750 Glu Ile Gly Glu Lys Gly Ile Asn Leu Ser Gly Gly Gln Lys Gln Arg 755760 765 Ile Ser Leu Ala Arg Ala Thr Tyr Gln Asn Leu Asp Ile Tyr Leu Leu770 775 780 Asp Asp Pro Leu Ser Ala Val Asp Ala His Val Gly Lys His IlePhe 785 790 795 800 Asn Lys Val Leu Gly Pro Asn Gly Leu Leu Lys Gly LysThr Arg Leu 805 810 815 Leu Val Thr His Ser Met His Phe Leu Pro Gln ValAsp Glu Ile Val 820 825 830 Val Leu Gly Asn Gly Thr Ile Val Glu Lys GlySer Tyr Ser Ala Leu 835 840 845 Leu Ala Lys Lys Gly Glu Phe Ala Lys AsnLeu Lys Thr Phe Leu Arg 850 855 860 His Thr Gly Pro Glu Glu Glu Ala ThrVal His Asp Gly Ser Glu Glu 865 870 875 880 Glu Asp Asp Asp Tyr Gly LeuIle Ser Ser Val Glu Glu Ile Pro Glu 885 890 895 Asp Ala Ala Ser Ile ThrMet Arg Arg Glu Asn Ser Phe Arg Arg Thr 900 905 910 Leu Ser Arg Ser SerArg Ser Asn Gly Arg His Leu Lys Ser Leu Arg 915 920 925 Asn Ser Leu LysThr Arg Asn Val Asn Ser Leu Lys Glu Asp Glu Glu 930 935 940 Leu Val LysGly Gln Lys Leu Ile Lys Lys Glu Phe Ile Glu Thr Gly 945 950 955 960 LysVal Lys Phe Ser Ile Tyr Leu Glu Tyr Leu Gln Ala Ile Gly Leu 965 970 975Phe Ser Ile Phe Phe Ile Ile Leu Ala Phe Val Met Asn Ser Val Ala 980 985990 Phe Ile Gly Ser Asn Leu Trp Leu Ser Ala Trp Thr Ser Asp Ser Lys 9951000 1005 Ile Phe Asn Ser Thr Asp Tyr Pro Ala Ser Gln Arg Asp Met Arg1010 1015 1020 Val Gly Val Tyr Gly Ala Leu Gly Leu Ala Gln Gly Ile PheVal 1025 1030 1035 Phe Ile Ala His Phe Trp Ser Ala Phe Gly Phe Val HisAla Ser 1040 1045 1050 Asn Ile Leu His Lys Gln Leu Leu Asn Asn Ile LeuArg Ala Pro 1055 1060 1065 Met Arg Phe Phe Asp Thr Thr Pro Thr Gly ArgIle Val Asn Arg 1070 1075 1080 Phe Ala Gly Asp Ile Ser Thr Val Asp AspThr Leu Pro Gln Ser 1085 1090 1095 Leu Arg Ser Trp Ile Thr Cys Phe LeuGly Ile Ile Ser Thr Leu 1100 1105 1110 Val Met Ile Cys Met Ala Thr ProVal Phe Thr Ile Ile Val Ile 1115 1120 1125 Pro Leu Gly Ile Ile Tyr ValSer Val Gln Met Phe Tyr Val Ser 1130 1135 1140 Thr Ser Arg Gln Leu ArgArg Leu Asp Ser Val Thr Arg Ser Pro 1145 1150 1155 Ile Tyr Ser His PheSer Glu Thr Val Ser Gly Leu Pro Val Ile 1160 1165 1170 Arg Ala Phe GluHis Gln Gln Arg Phe Leu Lys His Asn Glu Glu 1175 1180 1185 Arg Ile AspThr Asn Gln Lys Cys Val Phe Ser Trp Ile Thr Ser 1190 1195 1200 Asn ArgTrp Leu Ala Ile Arg Leu Glu Leu Val Gly Asn Leu Thr 1205 1210 1215 ValPhe Phe Ser Ala Leu Met Met Val Ile Tyr Arg Asp Thr Leu 1220 1225 1230Ser Gly Asp Thr Val Gly Phe Val Leu Ser Asn Ala Leu Asn Ile 1235 12401245 Thr Gln Thr Leu Asn Trp Leu Val Arg Met Thr Ser Glu Ile Glu 12501255 1260 Thr Asn Ile Val Ala Val Glu Arg Ile Thr Glu Tyr Thr Lys Val1265 1270 1275 Glu Asn Glu Ala Pro Trp Val Thr Asp Lys Arg Pro Pro ProAsp 1280 1285 1290 Trp Pro Ser Lys Gly Lys Ile Gln Phe Asn Asn Tyr GlnVal Arg 1295 1300 1305 Tyr Arg Pro Glu Leu Asp Leu Val Leu Arg Gly IleThr Cys Asp 1310 1315 1320 Ile Gly Ser Met Glu Lys Ile Gly Val Val GlyArg Thr Gly Ala 1325 1330 1335 Gly Lys Ser Ser Leu Thr Asn Cys Leu PheArg Ile Leu Glu Ala 1340 1345 1350 Ala Gly Gly Gln Ile Ile Ile Asp GlyVal Asp Ile Ala Ser Ile 1355 1360 1365 Gly Leu His Asp Leu Arg Glu LysLeu Thr Ile Ile Pro Gln Asp 1370 1375 1380 Pro Ile Leu Phe Ser Gly SerLeu Arg Met Asn Leu Asp Pro Phe 1385 1390 1395 Asn Asn Tyr Ser Asp GluGlu Ile Trp Lys Ala Leu Glu Leu Ala 1400 1405 1410 His Leu Lys Ser PheVal Ala Ser Leu Gln Leu Gly Leu Ser His 1415 1420 1425 Glu Val Thr GluAla Gly Gly Asn Leu Ser Ile Gly Gln Arg Gln 1430 1435 1440 Leu Leu CysLeu Gly Arg Ala Leu Leu Arg Lys Ser Lys Ile Leu 1445 1450 1455 Val LeuAsp Glu Ala Thr Ala Ala Val Asp Leu Glu Thr Asp Asn 1460 1465 1470 LeuIle Gln Thr Thr Ile Gln Asn Glu Phe Ala His Cys Thr Val 1475 1480 1485Ile Thr Ile Ala His Arg Leu His Thr Ile Met Asp Ser Asp Lys 1490 14951500 Val Met Val Leu Asp Asn Gly Lys Ile Ile Glu Tyr Gly Ser Pro 15051510 1515 Glu Glu Leu Leu Gln Ile Pro Gly Pro Phe Tyr Phe Met Ala Lys1520 1525 1530 Glu Ala Gly Ile Glu Asn Val Asn Ser Ala Ala Ala 1535 15401545 17 42 PRT Homo sapiens 17 Val Met Val Leu Asp Asn Gly Lys Ile IleGlu Tyr Gly Ser Pro Glu 1 5 10 15 Glu Leu Leu Gln Ile Pro Gly Pro PheTyr Phe Met Ala Lys Glu Ala 20 25 30 Gly Ile Glu Asn Val Asn Ser Thr LysPhe 35 40 18 42 PRT R. rattus 18 Ile Met Val Leu Asp Asn Gly Lys Ile ValGlu Tyr Gly Ser Pro Glu 1 5 10 15 Glu Leu Leu Ser Asn Arg Gly Ser PheTyr Leu Met Ala Lys Glu Ala 20 25 30 Gly Ile Glu Asn Val Asn His Thr GluLeu 35 40 19 43 PRT Oryctolagus 19 Ile Met Val Leu Asp Asn Gly Asn IleVal Glu Tyr Gly Ser Pro Glu 1 5 10 15 Glu Leu Leu Glu Ser Ala Gly ProPhe Ser Leu Met Ala Lys Glu Ser 20 25 30 Gly Ile Glu Asn Val Asn Asn ThrAla Phe Trp 35 40 20 25 PRT Homo sapiens 20 Val Val Asn Gly Arg Val LysHis Gly Thr His Ala Lys Gly Tyr Ser 1 5 10 15 Met Val Ser Val Ala GlyThr Lys Arg 20 25 21 36 PRT R. rattus 21 Ile Val Val Ile Gln Asn Gly GlnVal Lys Glu His Gly Thr His Gln 1 5 10 15 Gln Leu Leu Ala Gln Lys GlyIle Tyr Phe Ser Met Val Gln Ala Gly 20 25 30 Ala Lys Arg Ser 35 22 38PRT Homo sapiens 22 Ile Val Val Phe Gln Asn Gly Arg Val Lys Glu His GlyThr His Gln 1 5 10 15 Gln Leu Leu Ala Gln Lys Gly Ile Tyr Phe Ser MetVal Ser Val Gln 20 25 30 Ala Gly Thr Gln Asn Leu 35 23 34 PRT Homosapiens 23 Ile Val Leu Asp Lys Gly Glu Ile Gln Glu Tyr Gly Ala Pro SerAsp 1 5 10 15 Leu Leu Gln Gln Arg Gly Leu Phe Tyr Ser Met Ala Lys AspAla Gly 20 25 30 Leu Val 24 31 DNA Artificial Sequence Description ofArtificial Sequence oligonucleotide primer for amplifying human cMOAT 24agcgctagcg atgctggaga agttctgcaa c 31 25 36 DNA Artificial SequenceDescription of Artificial Sequence oligonucleotide primer for amplifyinghuman cMOAT 25 tacggtaccg gtgcgaattt tgtgctgttc acattc 36 26 25 DNAArtificial Sequence Description of Artificial Sequence oligonucleotideprimer for amplifying delta cMOAT (no gfp fusion) 26 gaatgtgaacagcacaaaat tcgcc 25 27 25 DNA Artificial Sequence Description ofArtificial Sequence oligonucleotide primer for amplifying T1543A K1544PF1545V 27 gaatgtgaac agcgcaccgg tcgcc 25 28 24 DNA Artificial SequenceDescription of Artificial Sequence oligonucleotide primer for amplifyingS1542A 28 gaatgtgaac gccacaaaat tcgc 24 29 24 DNA Artificial SequenceDescription of Artificial Sequence oligonucleotide primer for amplifyingT1543A 29 tgtgaacagc gcaaaattcg cacc 24 30 24 DNA Artificial SequenceDescription of Artificial Sequence oligonucleotide primer for amplifyingK1544A 30 gtgaacagca cagcattcgc accg 24 31 23 DNA Artificial SequenceDescription of Artificial Sequence oligonucleotide primer for amplifyingF1545A 31 cagcacaaaa gccgcaccgg tcg 23 32 30 DNA Artificial SequenceDescription of Artificial Sequence oligonucleotide primer for amplifyingT1543A K1544A F1545A 32 atgtgaacag cgcagcagcc gcaccggtcg 30 33 25 DNAArtificial Sequence Description of Artificial Sequence oligonucleotideprimer for amplifying delta cMOAT (for gfp fusion) 33 gaatgtgaacagctagcaga aggcc 25 34 28 DNA Artificial Sequence Description ofArtificial Sequence oligonucleotide primer for amplifying human MRP1 34gcggccgcgg atggcgctcc ggggcttc 28 35 34 DNA Artificial SequenceDescription of Artificial Sequence oligonucleotide primer for amplifyinghuman MRP1 35 tacggtaccg gtgccaccaa gccggcgtct ttgg 34 36 25 DNAArtificial Sequence Description of Artificial Sequence oligonucleotideprimer for amplifying non-tagged delta cMOAT 36 ggccttctgc tagctgttcacattc 25 37 7 PRT Homo sapiens 37 Asn Val Asn Ser Thr Lys Phe 1 5 38 7PRT Artificial Sequence Description of Artificial Sequence cMOAT T1543AK1544P F1545V mutant C-terminus 38 Asn Val Asn Ser Ala Pro Val 1 5 39 7PRT Artificial Sequence Description of Artificial Sequence cMOAT S1542Amutant C-terminus 39 Asn Val Asn Ala Thr Lys Phe 1 5 40 6 PRT ArtificialSequence Description of Artificial Sequence cMOAT T1543A mutantC-terminus 40 Val Asn Ser Ala Lys Phe 1 5 41 6 PRT Artificial SequenceDescription of Artificial Sequence cMOAT K1544A mutant C-terminus 41 ValAsn Ser Thr Ala Phe 1 5 42 4 PRT Artificial Sequence Description ofArtificial Sequence cMOAT F1545A mutant C-terminus 42 Ser Thr Lys Ala 143 6 PRT Artificial Sequence Description of Artificial Sequence cMOATT1543A K1544A F1545A mutant C-terminus 43 Val Asn Ser Ala Ala Ala 1 5 444 PRT Artificial Sequence Description of Artificial Sequence delta cMOATC-terminus 44 Asn Val Asn Ser 1 45 40 PRT Artificial SequenceDescription of Artificial Sequence HisP C-terminus 45 Val Ile Phe LeuHis Gln Gly Lys Ile Glu Glu Glu Gly Asp Pro Glu 1 5 10 15 Gln Val PheGly Asn Pro Gln Ser Pro Arg Leu Gln Gln Phe Leu Lys 20 25 30 Gly Ser LeuLys Lys Leu Glu His 35 40 46 42 PRT Mus musculus 46 Val Met Val Leu AspSer Gly Lys Ile Val Glu Tyr Gly Ser Pro Glu 1 5 10 15 Glu Leu Leu SerAsn Met Gly Pro Phe Tyr Leu Met Ala Lys Glu Ala 20 25 30 Gly Ile Glu SerVal Asn His Thr Glu Leu 35 40 47 82 PRT Homo sapiens 47 Ile Met Val LeuAsp Ser Gly Arg Leu Lys Glu Tyr Asp Glu Pro Tyr 1 5 10 15 Val Leu LeuGln Asn Lys Glu Ser Leu Phe Tyr Lys Met Val Gln Gln 20 25 30 Leu Gly LysAla Glu Ala Ala Ala Leu Thr Glu Thr Ala Lys Gln Val 35 40 45 Tyr Phe LysArg Asn Tyr Pro His Ile Gly His Thr Asp His Met Val 50 55 60 Thr Asn ThrSer Asn Gly Gln Pro Ser Thr Leu Thr Ile Phe Glu Thr 65 70 75 80 Ala Leu48 3825 DNA Homo sapiens CDS (1)..(3825) 48 atg gat ctt gag gcg gca aagaac gga aca gcc tgg cgc ccc acg agc 48 Met Asp Leu Glu Ala Ala Lys AsnGly Thr Ala Trp Arg Pro Thr Ser 1 5 10 15 gcg gag ggc gac ttt gaa ctgggc atc agc agc aaa caa aaa agg aaa 96 Ala Glu Gly Asp Phe Glu Leu GlyIle Ser Ser Lys Gln Lys Arg Lys 20 25 30 aaa acg aag aca gtg aaa atg attgga gta tta aca ttg ttt cga tac 144 Lys Thr Lys Thr Val Lys Met Ile GlyVal Leu Thr Leu Phe Arg Tyr 35 40 45 tcc gat tgg cag gat aaa ttg ttt atgtcg ctg ggt acc atc atg gcc 192 Ser Asp Trp Gln Asp Lys Leu Phe Met SerLeu Gly Thr Ile Met Ala 50 55 60 ata gct cac gga tca ggt ctc ccc ctc atgatg ata gta ttt gga gag 240 Ile Ala His Gly Ser Gly Leu Pro Leu Met MetIle Val Phe Gly Glu 65 70 75 80 atg act gac aaa ttt gtt gat act gca ggaaac ttc tcc ttt cca gtg 288 Met Thr Asp Lys Phe Val Asp Thr Ala Gly AsnPhe Ser Phe Pro Val 85 90 95 aac ttt tcc ttg tcg ctg cta aat cca ggc aaaatt ctg gaa gaa gaa 336 Asn Phe Ser Leu Ser Leu Leu Asn Pro Gly Lys IleLeu Glu Glu Glu 100 105 110 atg act aga tat gca tat tac tac tca gga ttgggt gct gga gtt ctt 384 Met Thr Arg Tyr Ala Tyr Tyr Tyr Ser Gly Leu GlyAla Gly Val Leu 115 120 125 gtt gct gcc tat ata caa gtt tca ttt tgg actttg gca gct ggt cga 432 Val Ala Ala Tyr Ile Gln Val Ser Phe Trp Thr LeuAla Ala Gly Arg 130 135 140 cag atc agg aaa att agg cag aag ttt ttt catgct att cta cga cag 480 Gln Ile Arg Lys Ile Arg Gln Lys Phe Phe His AlaIle Leu Arg Gln 145 150 155 160 gaa ata gga tgg ttt gac atc aac gac accact gaa ctc aat acg cgg 528 Glu Ile Gly Trp Phe Asp Ile Asn Asp Thr ThrGlu Leu Asn Thr Arg 165 170 175 cta aca gat gac atc tcc aaa atc agt gaagga att ggt gac aag gtt 576 Leu Thr Asp Asp Ile Ser Lys Ile Ser Glu GlyIle Gly Asp Lys Val 180 185 190 gga atg ttc ttt caa gca gta gcc acg tttttt gca gga ttc ata gtg 624 Gly Met Phe Phe Gln Ala Val Ala Thr Phe PheAla Gly Phe Ile Val 195 200 205 gga ttc atc aga gga tgg aag ctc acc cttgtg ata atg gcc atc agc 672 Gly Phe Ile Arg Gly Trp Lys Leu Thr Leu ValIle Met Ala Ile Ser 210 215 220 cct att cta gga ctc tct gca gcc gtt tgggca aag ata ctc tcg gca 720 Pro Ile Leu Gly Leu Ser Ala Ala Val Trp AlaLys Ile Leu Ser Ala 225 230 235 240 ttt agt gac aaa gaa cta gct gct tatgca aaa gca ggc gcc gtg gca 768 Phe Ser Asp Lys Glu Leu Ala Ala Tyr AlaLys Ala Gly Ala Val Ala 245 250 255 gaa gag gct ctg ggg gcc atc agg actgtg ata gct ttc ggg ggc cag 816 Glu Glu Ala Leu Gly Ala Ile Arg Thr ValIle Ala Phe Gly Gly Gln 260 265 270 aac aaa gag ctg gaa agg tat cag aaacat tta gaa aat gcc aaa gag 864 Asn Lys Glu Leu Glu Arg Tyr Gln Lys HisLeu Glu Asn Ala Lys Glu 275 280 285 att gga att aaa aaa gct att tca gcaaac att tcc atg ggt att gcc 912 Ile Gly Ile Lys Lys Ala Ile Ser Ala AsnIle Ser Met Gly Ile Ala 290 295 300 ttc ctg tta ata tat gca tca tat gcactg gcc ttc tgg tat gga tcc 960 Phe Leu Leu Ile Tyr Ala Ser Tyr Ala LeuAla Phe Trp Tyr Gly Ser 305 310 315 320 act cta gtc ata tca aaa gaa tatact att gga aat gca atg aca gtt 1008 Thr Leu Val Ile Ser Lys Glu Tyr ThrIle Gly Asn Ala Met Thr Val 325 330 335 ttt ttt tca atc cta att gga gctttc agt gtt ggc cag gct gcc cca 1056 Phe Phe Ser Ile Leu Ile Gly Ala PheSer Val Gly Gln Ala Ala Pro 340 345 350 tgt att gat gct ttt gcc aat gcaaga gga gca gca tat gtg atc ttt 1104 Cys Ile Asp Ala Phe Ala Asn Ala ArgGly Ala Ala Tyr Val Ile Phe 355 360 365 gat att att gat aat aat cct aaaatt gac agt ttt tca gag aga gga 1152 Asp Ile Ile Asp Asn Asn Pro Lys IleAsp Ser Phe Ser Glu Arg Gly 370 375 380 cac aaa cca gac agc atc aaa gggaat ttg gag ttc aat gat gtt cac 1200 His Lys Pro Asp Ser Ile Lys Gly AsnLeu Glu Phe Asn Asp Val His 385 390 395 400 ttt tct tac cct tct cga gctaac gtc aag atc ttg aag ggc ctc aac 1248 Phe Ser Tyr Pro Ser Arg Ala AsnVal Lys Ile Leu Lys Gly Leu Asn 405 410 415 ctg aag gtg cag agt ggg cagacg gtg gcc ctg gtt gga agt agt ggc 1296 Leu Lys Val Gln Ser Gly Gln ThrVal Ala Leu Val Gly Ser Ser Gly 420 425 430 tgt ggg aag agc aca acg gtccag ctg ata cag agg ctc tat gac cct 1344 Cys Gly Lys Ser Thr Thr Val GlnLeu Ile Gln Arg Leu Tyr Asp Pro 435 440 445 gat gag ggc aca att aac attgat ggg cag gat att agg aac ttt aat 1392 Asp Glu Gly Thr Ile Asn Ile AspGly Gln Asp Ile Arg Asn Phe Asn 450 455 460 gta aac tat ctg agg gaa atcatt ggt gtg gtg agt cag gag ccg gtg 1440 Val Asn Tyr Leu Arg Glu Ile IleGly Val Val Ser Gln Glu Pro Val 465 470 475 480 ctg ttt tcc acc aca attgct gaa aat att tgt tat ggc cgt gga aat 1488 Leu Phe Ser Thr Thr Ile AlaGlu Asn Ile Cys Tyr Gly Arg Gly Asn 485 490 495 gta acc atg gat gag ataaag aaa gct gtc aaa gag gcc aac gcc tat 1536 Val Thr Met Asp Glu Ile LysLys Ala Val Lys Glu Ala Asn Ala Tyr 500 505 510 gag ttt atc atg aaa ttacca cag aaa ttt gac acc ctg gtt gga gag 1584 Glu Phe Ile Met Lys Leu ProGln Lys Phe Asp Thr Leu Val Gly Glu 515 520 525 aga ggg gcc cag ctg agtggt ggg cag aag cag agg atc gcc att gca 1632 Arg Gly Ala Gln Leu Ser GlyGly Gln Lys Gln Arg Ile Ala Ile Ala 530 535 540 cgt gcc ctg gtt cgc aacccc aag atc ctt ctg ctg gat gag gcc acg 1680 Arg Ala Leu Val Arg Asn ProLys Ile Leu Leu Leu Asp Glu Ala Thr 545 550 555 560 tca gca ttg gac acagaa agt gaa gct gag gta cag gca gct ctg gat 1728 Ser Ala Leu Asp Thr GluSer Glu Ala Glu Val Gln Ala Ala Leu Asp 565 570 575 aag gcc aga gaa ggccgg acc acc att gtg ata gca cac cga ctg tct 1776 Lys Ala Arg Glu Gly ArgThr Thr Ile Val Ile Ala His Arg Leu Ser 580 585 590 acg gtc cga aat gcagat gtc atc gct ggg ttt gag gat gga gta att 1824 Thr Val Arg Asn Ala AspVal Ile Ala Gly Phe Glu Asp Gly Val Ile 595 600 605 gtg gag caa gga agccac agc gaa ctg atg aag aag gaa ggg gtg tac 1872 Val Glu Gln Gly Ser HisSer Glu Leu Met Lys Lys Glu Gly Val Tyr 610 615 620 ttc aaa ctt gtc aacatg cag aca tca gga agc cag atc cag tca gaa 1920 Phe Lys Leu Val Asn MetGln Thr Ser Gly Ser Gln Ile Gln Ser Glu 625 630 635 640 gaa ttt gaa ctaaat gat gaa aag gct gcc act aga atg gcc cca aat 1968 Glu Phe Glu Leu AsnAsp Glu Lys Ala Ala Thr Arg Met Ala Pro Asn 645 650 655 ggc tgg aaa tctcgc cta ttt agg cat tct act cag aaa aac ctt aaa 2016 Gly Trp Lys Ser ArgLeu Phe Arg His Ser Thr Gln Lys Asn Leu Lys 660 665 670 aat tca caa atgtgt cag aag agc ctt gat gtg gaa acc gat gga ctt 2064 Asn Ser Gln Met CysGln Lys Ser Leu Asp Val Glu Thr Asp Gly Leu 675 680 685 gaa gca aat gtgcca cca gtg tcc ttt ctg aag gtc ctg aaa ctg aat 2112 Glu Ala Asn Val ProPro Val Ser Phe Leu Lys Val Leu Lys Leu Asn 690 695 700 aaa aca gaa tggccc tac ttt gtc gtg gga aca gta tgt gcc att gcc 2160 Lys Thr Glu Trp ProTyr Phe Val Val Gly Thr Val Cys Ala Ile Ala 705 710 715 720 aat ggg gggctt cag ccg gca ttt tca gtc ata ttc tca gag atc ata 2208 Asn Gly Gly LeuGln Pro Ala Phe Ser Val Ile Phe Ser Glu Ile Ile 725 730 735 gcg att tttgga cca ggc gat gat gca gtg aag cag cag aag tgc aac 2256 Ala Ile Phe GlyPro Gly Asp Asp Ala Val Lys Gln Gln Lys Cys Asn 740 745 750 ata ttc tctttg att ttc tta ttt ctg gga att att tct ttt ttt act 2304 Ile Phe Ser LeuIle Phe Leu Phe Leu Gly Ile Ile Ser Phe Phe Thr 755 760 765 ttc ttc cttcag ggt ttc acg ttt ggg aaa gct ggc gag atc ctc acc 2352 Phe Phe Leu GlnGly Phe Thr Phe Gly Lys Ala Gly Glu Ile Leu Thr 770 775 780 aga aga ctgcgg tca atg gct ttt aaa gca atg cta aga cag gac atg 2400 Arg Arg Leu ArgSer Met Ala Phe Lys Ala Met Leu Arg Gln Asp Met 785 790 795 800 agc tggttt gat gac cat aaa aac agt act ggt gca ctt tct aca aga 2448 Ser Trp PheAsp Asp His Lys Asn Ser Thr Gly Ala Leu Ser Thr Arg 805 810 815 ctt gccaca gat gct gcc caa gtc caa gga gcc aca gga acc agg ttg 2496 Leu Ala ThrAsp Ala Ala Gln Val Gln Gly Ala Thr Gly Thr Arg Leu 820 825 830 gct ttaatt gca cag aat ata gct aac ctt gga act ggt att atc ata 2544 Ala Leu IleAla Gln Asn Ile Ala Asn Leu Gly Thr Gly Ile Ile Ile 835 840 845 tca tttatc tac ggt tgg cag tta acc cta ttg cta tta gca gtt gtt 2592 Ser Phe IleTyr Gly Trp Gln Leu Thr Leu Leu Leu Leu Ala Val Val 850 855 860 cca attatt gct gtg tca gga att gtt gaa atg aaa ttg ttg gct gga 2640 Pro Ile IleAla Val Ser Gly Ile Val Glu Met Lys Leu Leu Ala Gly 865 870 875 880 aatgcc aaa aga gat aaa aaa gaa ctg gaa gct gct gga aag att gca 2688 Asn AlaLys Arg Asp Lys Lys Glu Leu Glu Ala Ala Gly Lys Ile Ala 885 890 895 acagag gca ata gaa aat att agg aca gtt gtg tct ttg acc cag gaa 2736 Thr GluAla Ile Glu Asn Ile Arg Thr Val Val Ser Leu Thr Gln Glu 900 905 910 agaaaa ttt gaa tca atg tat gtt gaa aaa ttg tat gga cct tac agg 2784 Arg LysPhe Glu Ser Met Tyr Val Glu Lys Leu Tyr Gly Pro Tyr Arg 915 920 925 aattct gtg cag aag gca cac atc tat gga att act ttt agt atc tca 2832 Asn SerVal Gln Lys Ala His Ile Tyr Gly Ile Thr Phe Ser Ile Ser 930 935 940 caagca ttt atg tat ttt tcc tat gcc ggt tgt ttt cga ttt ggt gca 2880 Gln AlaPhe Met Tyr Phe Ser Tyr Ala Gly Cys Phe Arg Phe Gly Ala 945 950 955 960tat ctc att gtg aat gga cat atg cgc ttc aga gat gtt att ctg gtg 2928 TyrLeu Ile Val Asn Gly His Met Arg Phe Arg Asp Val Ile Leu Val 965 970 975ttt tct gca att gta ttt ggt gca gtg gct cta gga cat gcc agt tca 2976 PheSer Ala Ile Val Phe Gly Ala Val Ala Leu Gly His Ala Ser Ser 980 985 990ttt gct cca gac tat gct aaa gct aag ctg tct gca gcc cac tta ttc 3024 PheAla Pro Asp Tyr Ala Lys Ala Lys Leu Ser Ala Ala His Leu Phe 995 10001005 atg ctg ttt gaa aga caa cct ctg att gac agc tac agt gaa gag 3069Met Leu Phe Glu Arg Gln Pro Leu Ile Asp Ser Tyr Ser Glu Glu 1010 10151020 ggg ctg aag cct gat aaa ttt gaa gga aat ata aca ttt aat gaa 3114Gly Leu Lys Pro Asp Lys Phe Glu Gly Asn Ile Thr Phe Asn Glu 1025 10301035 gtc gtg ttc aac tat ccc acc cga gca aac gtg cca gtg ctt cag 3159Val Val Phe Asn Tyr Pro Thr Arg Ala Asn Val Pro Val Leu Gln 1040 10451050 ggg ctg agc ctg gag gtg aag aaa ggc cag aca cta gcc ctg gtg 3204Gly Leu Ser Leu Glu Val Lys Lys Gly Gln Thr Leu Ala Leu Val 1055 10601065 ggc agc agt ggc tgt ggg aag agc acg gtg gtc cag ctc ctg gag 3249Gly Ser Ser Gly Cys Gly Lys Ser Thr Val Val Gln Leu Leu Glu 1070 10751080 cgg ttc tac gac ccc ttg gcg ggg aca gtg ctt ctc gat ggt caa 3294Arg Phe Tyr Asp Pro Leu Ala Gly Thr Val Leu Leu Asp Gly Gln 1085 10901095 gaa gca aag aaa ctc aat gtc cag tgg ctc aga gct caa ctc gga 3339Glu Ala Lys Lys Leu Asn Val Gln Trp Leu Arg Ala Gln Leu Gly 1100 11051110 atc gtg tct cag gag cct atc cta ttt gac tgc agc att gcc gag 3384Ile Val Ser Gln Glu Pro Ile Leu Phe Asp Cys Ser Ile Ala Glu 1115 11201125 aat att gcc tat gga gac aac agc cgg gtt gta tca cag gat gaa 3429Asn Ile Ala Tyr Gly Asp Asn Ser Arg Val Val Ser Gln Asp Glu 1130 11351140 att gtg agt gca gcc aaa gct gcc aac ata cat cct ttc atc gag 3474Ile Val Ser Ala Ala Lys Ala Ala Asn Ile His Pro Phe Ile Glu 1145 11501155 acg tta ccc cac aaa tat gaa aca aga gtg gga gat aag ggg act 3519Thr Leu Pro His Lys Tyr Glu Thr Arg Val Gly Asp Lys Gly Thr 1160 11651170 cag ctc tca gga ggt caa aaa cag agg att gct att gcc cga gcc 3564Gln Leu Ser Gly Gly Gln Lys Gln Arg Ile Ala Ile Ala Arg Ala 1175 11801185 ctc atc aga caa cct caa atc ctc ctg ttg gat gaa gct aca tca 3609Leu Ile Arg Gln Pro Gln Ile Leu Leu Leu Asp Glu Ala Thr Ser 1190 11951200 gct ctg gat act gaa agt gaa aag gtt gtc caa gaa gcc ctg gac 3654Ala Leu Asp Thr Glu Ser Glu Lys Val Val Gln Glu Ala Leu Asp 1205 12101215 aaa gcc aga gaa ggc cgc acc tgc att gtg att gct cac cgc ctg 3699Lys Ala Arg Glu Gly Arg Thr Cys Ile Val Ile Ala His Arg Leu 1220 12251230 tcc acc atc cag aat gca gac tta ata gtg gtg ttt cag aat ggg 3744Ser Thr Ile Gln Asn Ala Asp Leu Ile Val Val Phe Gln Asn Gly 1235 12401245 aga gtc aag gag cat ggc acg cat cag cag ctg ctg gca cag aaa 3789Arg Val Lys Glu His Gly Thr His Gln Gln Leu Leu Ala Gln Lys 1250 12551260 ggc atc tat ttt tca atg gtc agt gtc cag gct ggg 3825 Gly Ile TyrPhe Ser Met Val Ser Val Gln Ala Gly 1265 1270 1275 49 1275 PRT Homosapiens 49 Met Asp Leu Glu Ala Ala Lys Asn Gly Thr Ala Trp Arg Pro ThrSer 1 5 10 15 Ala Glu Gly Asp Phe Glu Leu Gly Ile Ser Ser Lys Gln LysArg Lys 20 25 30 Lys Thr Lys Thr Val Lys Met Ile Gly Val Leu Thr Leu PheArg Tyr 35 40 45 Ser Asp Trp Gln Asp Lys Leu Phe Met Ser Leu Gly Thr IleMet Ala 50 55 60 Ile Ala His Gly Ser Gly Leu Pro Leu Met Met Ile Val PheGly Glu 65 70 75 80 Met Thr Asp Lys Phe Val Asp Thr Ala Gly Asn Phe SerPhe Pro Val 85 90 95 Asn Phe Ser Leu Ser Leu Leu Asn Pro Gly Lys Ile LeuGlu Glu Glu 100 105 110 Met Thr Arg Tyr Ala Tyr Tyr Tyr Ser Gly Leu GlyAla Gly Val Leu 115 120 125 Val Ala Ala Tyr Ile Gln Val Ser Phe Trp ThrLeu Ala Ala Gly Arg 130 135 140 Gln Ile Arg Lys Ile Arg Gln Lys Phe PheHis Ala Ile Leu Arg Gln 145 150 155 160 Glu Ile Gly Trp Phe Asp Ile AsnAsp Thr Thr Glu Leu Asn Thr Arg 165 170 175 Leu Thr Asp Asp Ile Ser LysIle Ser Glu Gly Ile Gly Asp Lys Val 180 185 190 Gly Met Phe Phe Gln AlaVal Ala Thr Phe Phe Ala Gly Phe Ile Val 195 200 205 Gly Phe Ile Arg GlyTrp Lys Leu Thr Leu Val Ile Met Ala Ile Ser 210 215 220 Pro Ile Leu GlyLeu Ser Ala Ala Val Trp Ala Lys Ile Leu Ser Ala 225 230 235 240 Phe SerAsp Lys Glu Leu Ala Ala Tyr Ala Lys Ala Gly Ala Val Ala 245 250 255 GluGlu Ala Leu Gly Ala Ile Arg Thr Val Ile Ala Phe Gly Gly Gln 260 265 270Asn Lys Glu Leu Glu Arg Tyr Gln Lys His Leu Glu Asn Ala Lys Glu 275 280285 Ile Gly Ile Lys Lys Ala Ile Ser Ala Asn Ile Ser Met Gly Ile Ala 290295 300 Phe Leu Leu Ile Tyr Ala Ser Tyr Ala Leu Ala Phe Trp Tyr Gly Ser305 310 315 320 Thr Leu Val Ile Ser Lys Glu Tyr Thr Ile Gly Asn Ala MetThr Val 325 330 335 Phe Phe Ser Ile Leu Ile Gly Ala Phe Ser Val Gly GlnAla Ala Pro 340 345 350 Cys Ile Asp Ala Phe Ala Asn Ala Arg Gly Ala AlaTyr Val Ile Phe 355 360 365 Asp Ile Ile Asp Asn Asn Pro Lys Ile Asp SerPhe Ser Glu Arg Gly 370 375 380 His Lys Pro Asp Ser Ile Lys Gly Asn LeuGlu Phe Asn Asp Val His 385 390 395 400 Phe Ser Tyr Pro Ser Arg Ala AsnVal Lys Ile Leu Lys Gly Leu Asn 405 410 415 Leu Lys Val Gln Ser Gly GlnThr Val Ala Leu Val Gly Ser Ser Gly 420 425 430 Cys Gly Lys Ser Thr ThrVal Gln Leu Ile Gln Arg Leu Tyr Asp Pro 435 440 445 Asp Glu Gly Thr IleAsn Ile Asp Gly Gln Asp Ile Arg Asn Phe Asn 450 455 460 Val Asn Tyr LeuArg Glu Ile Ile Gly Val Val Ser Gln Glu Pro Val 465 470 475 480 Leu PheSer Thr Thr Ile Ala Glu Asn Ile Cys Tyr Gly Arg Gly Asn 485 490 495 ValThr Met Asp Glu Ile Lys Lys Ala Val Lys Glu Ala Asn Ala Tyr 500 505 510Glu Phe Ile Met Lys Leu Pro Gln Lys Phe Asp Thr Leu Val Gly Glu 515 520525 Arg Gly Ala Gln Leu Ser Gly Gly Gln Lys Gln Arg Ile Ala Ile Ala 530535 540 Arg Ala Leu Val Arg Asn Pro Lys Ile Leu Leu Leu Asp Glu Ala Thr545 550 555 560 Ser Ala Leu Asp Thr Glu Ser Glu Ala Glu Val Gln Ala AlaLeu Asp 565 570 575 Lys Ala Arg Glu Gly Arg Thr Thr Ile Val Ile Ala HisArg Leu Ser 580 585 590 Thr Val Arg Asn Ala Asp Val Ile Ala Gly Phe GluAsp Gly Val Ile 595 600 605 Val Glu Gln Gly Ser His Ser Glu Leu Met LysLys Glu Gly Val Tyr 610 615 620 Phe Lys Leu Val Asn Met Gln Thr Ser GlySer Gln Ile Gln Ser Glu 625 630 635 640 Glu Phe Glu Leu Asn Asp Glu LysAla Ala Thr Arg Met Ala Pro Asn 645 650 655 Gly Trp Lys Ser Arg Leu PheArg His Ser Thr Gln Lys Asn Leu Lys 660 665 670 Asn Ser Gln Met Cys GlnLys Ser Leu Asp Val Glu Thr Asp Gly Leu 675 680 685 Glu Ala Asn Val ProPro Val Ser Phe Leu Lys Val Leu Lys Leu Asn 690 695 700 Lys Thr Glu TrpPro Tyr Phe Val Val Gly Thr Val Cys Ala Ile Ala 705 710 715 720 Asn GlyGly Leu Gln Pro Ala Phe Ser Val Ile Phe Ser Glu Ile Ile 725 730 735 AlaIle Phe Gly Pro Gly Asp Asp Ala Val Lys Gln Gln Lys Cys Asn 740 745 750Ile Phe Ser Leu Ile Phe Leu Phe Leu Gly Ile Ile Ser Phe Phe Thr 755 760765 Phe Phe Leu Gln Gly Phe Thr Phe Gly Lys Ala Gly Glu Ile Leu Thr 770775 780 Arg Arg Leu Arg Ser Met Ala Phe Lys Ala Met Leu Arg Gln Asp Met785 790 795 800 Ser Trp Phe Asp Asp His Lys Asn Ser Thr Gly Ala Leu SerThr Arg 805 810 815 Leu Ala Thr Asp Ala Ala Gln Val Gln Gly Ala Thr GlyThr Arg Leu 820 825 830 Ala Leu Ile Ala Gln Asn Ile Ala Asn Leu Gly ThrGly Ile Ile Ile 835 840 845 Ser Phe Ile Tyr Gly Trp Gln Leu Thr Leu LeuLeu Leu Ala Val Val 850 855 860 Pro Ile Ile Ala Val Ser Gly Ile Val GluMet Lys Leu Leu Ala Gly 865 870 875 880 Asn Ala Lys Arg Asp Lys Lys GluLeu Glu Ala Ala Gly Lys Ile Ala 885 890 895 Thr Glu Ala Ile Glu Asn IleArg Thr Val Val Ser Leu Thr Gln Glu 900 905 910 Arg Lys Phe Glu Ser MetTyr Val Glu Lys Leu Tyr Gly Pro Tyr Arg 915 920 925 Asn Ser Val Gln LysAla His Ile Tyr Gly Ile Thr Phe Ser Ile Ser 930 935 940 Gln Ala Phe MetTyr Phe Ser Tyr Ala Gly Cys Phe Arg Phe Gly Ala 945 950 955 960 Tyr LeuIle Val Asn Gly His Met Arg Phe Arg Asp Val Ile Leu Val 965 970 975 PheSer Ala Ile Val Phe Gly Ala Val Ala Leu Gly His Ala Ser Ser 980 985 990Phe Ala Pro Asp Tyr Ala Lys Ala Lys Leu Ser Ala Ala His Leu Phe 995 10001005 Met Leu Phe Glu Arg Gln Pro Leu Ile Asp Ser Tyr Ser Glu Glu 10101015 1020 Gly Leu Lys Pro Asp Lys Phe Glu Gly Asn Ile Thr Phe Asn Glu1025 1030 1035 Val Val Phe Asn Tyr Pro Thr Arg Ala Asn Val Pro Val LeuGln 1040 1045 1050 Gly Leu Ser Leu Glu Val Lys Lys Gly Gln Thr Leu AlaLeu Val 1055 1060 1065 Gly Ser Ser Gly Cys Gly Lys Ser Thr Val Val GlnLeu Leu Glu 1070 1075 1080 Arg Phe Tyr Asp Pro Leu Ala Gly Thr Val LeuLeu Asp Gly Gln 1085 1090 1095 Glu Ala Lys Lys Leu Asn Val Gln Trp LeuArg Ala Gln Leu Gly 1100 1105 1110 Ile Val Ser Gln Glu Pro Ile Leu PheAsp Cys Ser Ile Ala Glu 1115 1120 1125 Asn Ile Ala Tyr Gly Asp Asn SerArg Val Val Ser Gln Asp Glu 1130 1135 1140 Ile Val Ser Ala Ala Lys AlaAla Asn Ile His Pro Phe Ile Glu 1145 1150 1155 Thr Leu Pro His Lys TyrGlu Thr Arg Val Gly Asp Lys Gly Thr 1160 1165 1170 Gln Leu Ser Gly GlyGln Lys Gln Arg Ile Ala Ile Ala Arg Ala 1175 1180 1185 Leu Ile Arg GlnPro Gln Ile Leu Leu Leu Asp Glu Ala Thr Ser 1190 1195 1200 Ala Leu AspThr Glu Ser Glu Lys Val Val Gln Glu Ala Leu Asp 1205 1210 1215 Lys AlaArg Glu Gly Arg Thr Cys Ile Val Ile Ala His Arg Leu 1220 1225 1230 SerThr Ile Gln Asn Ala Asp Leu Ile Val Val Phe Gln Asn Gly 1235 1240 1245Arg Val Lys Glu His Gly Thr His Gln Gln Leu Leu Ala Gln Lys 1250 12551260 Gly Ile Tyr Phe Ser Met Val Ser Val Gln Ala Gly 1265 1270 1275 503966 DNA Homo sapiens CDS (1)..(3966) 50 atg ctg ccc gtg tac cag gag gtgaag ccc aac ccg ctg cag gac gcg 48 Met Leu Pro Val Tyr Gln Glu Val LysPro Asn Pro Leu Gln Asp Ala 1 5 10 15 aac ctc tgc tca cgc gtg ttc ttctgg tgg ctc aat ccc ttg ttt aaa 96 Asn Leu Cys Ser Arg Val Phe Phe TrpTrp Leu Asn Pro Leu Phe Lys 20 25 30 att ggc cat aaa cgg aga tta gag gaagat gat atg tat tca gtg ctg 144 Ile Gly His Lys Arg Arg Leu Glu Glu AspAsp Met Tyr Ser Val Leu 35 40 45 cca gaa gac cgc tca cag cac ctt gga gaggag ttg caa ggg ttc tgg 192 Pro Glu Asp Arg Ser Gln His Leu Gly Glu GluLeu Gln Gly Phe Trp 50 55 60 gat aaa gaa gtt tta aga gct gag aat gac gcacag aag cct tct tta 240 Asp Lys Glu Val Leu Arg Ala Glu Asn Asp Ala GlnLys Pro Ser Leu 65 70 75 80 aca aga gca atc ata aag tgt tac tgg aaa tcttat tta gtt ttg gga 288 Thr Arg Ala Ile Ile Lys Cys Tyr Trp Lys Ser TyrLeu Val Leu Gly 85 90 95 att ttt acg tta att gag gaa agt gcc aaa gta atccag ccc ata ttt 336 Ile Phe Thr Leu Ile Glu Glu Ser Ala Lys Val Ile GlnPro Ile Phe 100 105 110 ttg gga aaa att att aat tat ttt gaa aat tat gatccc atg gat tct 384 Leu Gly Lys Ile Ile Asn Tyr Phe Glu Asn Tyr Asp ProMet Asp Ser 115 120 125 gtg gct ttg aac aca gcg tac gcc tat gcc acg gtgctg act ttt tgc 432 Val Ala Leu Asn Thr Ala Tyr Ala Tyr Ala Thr Val LeuThr Phe Cys 130 135 140 acg ctc att ttg gct ata ctg cat cac tta tat ttttat cac gtt cag 480 Thr Leu Ile Leu Ala Ile Leu His His Leu Tyr Phe TyrHis Val Gln 145 150 155 160 tgt gct ggg atg agg tta cga gta gcc atg tgccat atg att tat cgg 528 Cys Ala Gly Met Arg Leu Arg Val Ala Met Cys HisMet Ile Tyr Arg 165 170 175 aag gca ctt cgt ctt agt aac atg gcc atg gggaag aca acc aca ggc 576 Lys Ala Leu Arg Leu Ser Asn Met Ala Met Gly LysThr Thr Thr Gly 180 185 190 cag ata gtc aat ctg ctg tcc aat gat gtg aacaag ttt gat cag gtg 624 Gln Ile Val Asn Leu Leu Ser Asn Asp Val Asn LysPhe Asp Gln Val 195 200 205 aca gtg ttc tta cac ttc ctg tgg gca gga ccactg cag gcg att gca 672 Thr Val Phe Leu His Phe Leu Trp Ala Gly Pro LeuGln Ala Ile Ala 210 215 220 gtg act gcc cta ctc tgg atg gag ata gga atatcg tgc ctt gct ggg 720 Val Thr Ala Leu Leu Trp Met Glu Ile Gly Ile SerCys Leu Ala Gly 225 230 235 240 atg gca gtt cta atc att ctc ctg ccc ttgcaa agc tgt ttt ggg aag 768 Met Ala Val Leu Ile Ile Leu Leu Pro Leu GlnSer Cys Phe Gly Lys 245 250 255 ttg ttc tca tca ctg agg agt aaa act gcaact ttc acg gat gcc agg 816 Leu Phe Ser Ser Leu Arg Ser Lys Thr Ala ThrPhe Thr Asp Ala Arg 260 265 270 atc agg acc atg aat gaa gtt ata act ggtata agg ata ata aaa atg 864 Ile Arg Thr Met Asn Glu Val Ile Thr Gly IleArg Ile Ile Lys Met 275 280 285 tac gcc tgg gaa aag tca ttt tca aat cttatt acc aat ttg aga aag 912 Tyr Ala Trp Glu Lys Ser Phe Ser Asn Leu IleThr Asn Leu Arg Lys 290 295 300 aag gag att tcc aag att ctg aga agt tcctgc ctc aga ggg atg aat 960 Lys Glu Ile Ser Lys Ile Leu Arg Ser Ser CysLeu Arg Gly Met Asn 305 310 315 320 ttg gct tca ttt ttc agt gca agc aaaatc atc gtg ttt gtg acc ttc 1008 Leu Ala Ser Phe Phe Ser Ala Ser Lys IleIle Val Phe Val Thr Phe 325 330 335 acc acc tac gtg ctc ctc ggc agt gtgatc aca gcc agc cgc gtg ttc 1056 Thr Thr Tyr Val Leu Leu Gly Ser Val IleThr Ala Ser Arg Val Phe 340 345 350 gtg gca gtg acg ctg tat ggg gct gtgcgg ctg acg gtt acc ctc ttc 1104 Val Ala Val Thr Leu Tyr Gly Ala Val ArgLeu Thr Val Thr Leu Phe 355 360 365 ttc ccc tca gcc att gag agg gtg tcagag gca atc gtc agc atc cga 1152 Phe Pro Ser Ala Ile Glu Arg Val Ser GluAla Ile Val Ser Ile Arg 370 375 380 aga atc cag acc ttt ttg cta ctt gatgag ata tca cag cgc aac cgt 1200 Arg Ile Gln Thr Phe Leu Leu Leu Asp GluIle Ser Gln Arg Asn Arg 385 390 395 400 cag ctg ccg tca gat ggt aaa aagatg gtg cat gtg cag gat ttt act 1248 Gln Leu Pro Ser Asp Gly Lys Lys MetVal His Val Gln Asp Phe Thr 405 410 415 gct ttt tgg gat aag gca tca gagacc cca act cta caa ggc ctt tcc 1296 Ala Phe Trp Asp Lys Ala Ser Glu ThrPro Thr Leu Gln Gly Leu Ser 420 425 430 ttt act gtc aga cct ggc gaa ttgtta gct gtg gtc ggc ccc gtg gga 1344 Phe Thr Val Arg Pro Gly Glu Leu LeuAla Val Val Gly Pro Val Gly 435 440 445 gca ggg aag tca tca ctg tta agtgcc gtg ctc ggg gaa ttg gcc cca 1392 Ala Gly Lys Ser Ser Leu Leu Ser AlaVal Leu Gly Glu Leu Ala Pro 450 455 460 agt cac ggg ctg gtc agc gtg catgga aga att gcc tat gtg tct cag 1440 Ser His Gly Leu Val Ser Val His GlyArg Ile Ala Tyr Val Ser Gln 465 470 475 480 cag ccc tgg gtg ttc tcg ggaact ctg agg agt aat att tta ttt ggg 1488 Gln Pro Trp Val Phe Ser Gly ThrLeu Arg Ser Asn Ile Leu Phe Gly 485 490 495 aag aaa tac gaa aag gaa cgatat gaa aaa gtc ata aag gct tgt gct 1536 Lys Lys Tyr Glu Lys Glu Arg TyrGlu Lys Val Ile Lys Ala Cys Ala 500 505 510 ctg aaa aag gat tta cag ctgttg gag gat ggt gat ctg act gtg ata 1584 Leu Lys Lys Asp Leu Gln Leu LeuGlu Asp Gly Asp Leu Thr Val Ile 515 520 525 gga gat cgg gga acc acg ctgagt gga ggg cag aaa gca cgg gta aac 1632 Gly Asp Arg Gly Thr Thr Leu SerGly Gly Gln Lys Ala Arg Val Asn 530 535 540 ctt gca aga gca gtg tat caagat gct gac atc tat ctc ctg gac gat 1680 Leu Ala Arg Ala Val Tyr Gln AspAla Asp Ile Tyr Leu Leu Asp Asp 545 550 555 560 cct ctc agt gca gta gatgcg gaa gtt agc aga cac ttg ttc gaa ctg 1728 Pro Leu Ser Ala Val Asp AlaGlu Val Ser Arg His Leu Phe Glu Leu 565 570 575 tgt att tgt caa att ttgcat gag aag atc aca att tta gtg act cat 1776 Cys Ile Cys Gln Ile Leu HisGlu Lys Ile Thr Ile Leu Val Thr His 580 585 590 cag ttg cag tac ctc aaagct gca agt cag att ctg ata ttg aaa gat 1824 Gln Leu Gln Tyr Leu Lys AlaAla Ser Gln Ile Leu Ile Leu Lys Asp 595 600 605 ggt aaa atg gtg cag aagggg act tac act gag ttc cta aaa tct ggt 1872 Gly Lys Met Val Gln Lys GlyThr Tyr Thr Glu Phe Leu Lys Ser Gly 610 615 620 ata gat ttt ggc tcc ctttta aag aag gat aat gag gaa agt gaa caa 1920 Ile Asp Phe Gly Ser Leu LeuLys Lys Asp Asn Glu Glu Ser Glu Gln 625 630 635 640 cct cca gtt cca ggaact ccc aca cta agg aat cgt acc ttc tca gag 1968 Pro Pro Val Pro Gly ThrPro Thr Leu Arg Asn Arg Thr Phe Ser Glu 645 650 655 tct tcg gtt tgg tctcaa caa tct tct aga ccc tcc ttg aaa gat ggt 2016 Ser Ser Val Trp Ser GlnGln Ser Ser Arg Pro Ser Leu Lys Asp Gly 660 665 670 gct ctg gag agc caagat aca gag aat gtc cca gtt aca cta tca gag 2064 Ala Leu Glu Ser Gln AspThr Glu Asn Val Pro Val Thr Leu Ser Glu 675 680 685 gag aac cgt tct gaagga aaa gtt ggt ttt cag gcc tat aag aat tac 2112 Glu Asn Arg Ser Glu GlyLys Val Gly Phe Gln Ala Tyr Lys Asn Tyr 690 695 700 ttc aga gct ggt gctcac tgg att gtc ttc att ttc ctt att ctc cta 2160 Phe Arg Ala Gly Ala HisTrp Ile Val Phe Ile Phe Leu Ile Leu Leu 705 710 715 720 aac act gca gctcag gtt gcc tat gtg ctt caa gat tgg tgg ctt tca 2208 Asn Thr Ala Ala GlnVal Ala Tyr Val Leu Gln Asp Trp Trp Leu Ser 725 730 735 tac tgg gca aacaaa caa agt atg cta aat gtc act gta aat gga gga 2256 Tyr Trp Ala Asn LysGln Ser Met Leu Asn Val Thr Val Asn Gly Gly 740 745 750 gga aat gta accgag aag cta gat ctt aac tgg tac tta gga att tat 2304 Gly Asn Val Thr GluLys Leu Asp Leu Asn Trp Tyr Leu Gly Ile Tyr 755 760 765 tca ggt tta actgta gct acc gtt ctt ttt ggc ata gca aga tct cta 2352 Ser Gly Leu Thr ValAla Thr Val Leu Phe Gly Ile Ala Arg Ser Leu 770 775 780 ttg gta ttc tacgtc ctt gtt aac tct tca caa act ttg cac aac aaa 2400 Leu Val Phe Tyr ValLeu Val Asn Ser Ser Gln Thr Leu His Asn Lys 785 790 795 800 atg ttt gagtca att ctg aaa gct ccg gta tta ttc ttt gat aga aat 2448 Met Phe Glu SerIle Leu Lys Ala Pro Val Leu Phe Phe Asp Arg Asn 805 810 815 cca ata ggaaga att tta aat cgt ttc tcc aaa gac att gga cac ttg 2496 Pro Ile Gly ArgIle Leu Asn Arg Phe Ser Lys Asp Ile Gly His Leu 820 825 830 gat gat ttgctg ccg ctg acg ttt tta gat ttc atc cag aca ttg cta 2544 Asp Asp Leu LeuPro Leu Thr Phe Leu Asp Phe Ile Gln Thr Leu Leu 835 840 845 caa gtg gttggt gtg gtc tct gtg gct gtg gcc gtg att cct tgg atc 2592 Gln Val Val GlyVal Val Ser Val Ala Val Ala Val Ile Pro Trp Ile 850 855 860 gca ata cccttg gtt ccc ctt gga atc att ttc att ttt ctt cgg cga 2640 Ala Ile Pro LeuVal Pro Leu Gly Ile Ile Phe Ile Phe Leu Arg Arg 865 870 875 880 tat tttttg gaa acg tca aga gat gtg aag cgc ctg gaa tct aca act 2688 Tyr Phe LeuGlu Thr Ser Arg Asp Val Lys Arg Leu Glu Ser Thr Thr 885 890 895 cgg agtcca gtg ttt tcc cac tta tca tct tct ctc cag ggg ctc tgg 2736 Arg Ser ProVal Phe Ser His Leu Ser Ser Ser Leu Gln Gly Leu Trp 900 905 910 acc atccgg gca tac aaa gca gaa gag agg tgt cag gaa ctg ttt gat 2784 Thr Ile ArgAla Tyr Lys Ala Glu Glu Arg Cys Gln Glu Leu Phe Asp 915 920 925 gca caccag gat tta cat tca gag gct tgg ttc ttg ttt ttg aca acg 2832 Ala His GlnAsp Leu His Ser Glu Ala Trp Phe Leu Phe Leu Thr Thr 930 935 940 tcc cgctgg ttt gcc gtc cgt ctg gat gcc atc tgt gcc atg ttt gtc 2880 Ser Arg TrpPhe Ala Val Arg Leu Asp Ala Ile Cys Ala Met Phe Val 945 950 955 960 atcatc gtt gcc ttt ggg tcc ctg att ctg gca aaa act ctg gat gcc 2928 Ile IleVal Ala Phe Gly Ser Leu Ile Leu Ala Lys Thr Leu Asp Ala 965 970 975 gggcag gtt ggt ttg gca ctg tcc tat gcc ctc acg ctc atg ggg atg 2976 Gly GlnVal Gly Leu Ala Leu Ser Tyr Ala Leu Thr Leu Met Gly Met 980 985 990 tttcag tgg tgt gtt cga caa agt gct gaa gtt gag aat atg atg atc 3024 Phe GlnTrp Cys Val Arg Gln Ser Ala Glu Val Glu Asn Met Met Ile 995 1000 1005tca gta gaa agg gtc att gaa tac aca gac ctt gaa aaa gaa gca 3069 Ser ValGlu Arg Val Ile Glu Tyr Thr Asp Leu Glu Lys Glu Ala 1010 1015 1020 ccttgg gaa tat cag aaa cgc cca cca cca gcc tgg ccc cat gaa 3114 Pro Trp GluTyr Gln Lys Arg Pro Pro Pro Ala Trp Pro His Glu 1025 1030 1035 gga gtgata atc ttt gac aat gtg aac ttc atg tac agt cca ggt 3159 Gly Val Ile IlePhe Asp Asn Val Asn Phe Met Tyr Ser Pro Gly 1040 1045 1050 ggg cct ctggta ctg aag cat ctg aca gca ctc att aaa tca caa 3204 Gly Pro Leu Val LeuLys His Leu Thr Ala Leu Ile Lys Ser Gln 1055 1060 1065 gaa aag gtt ggcatt gtg gga aga acc gga gct gga aaa agt tcc 3249 Glu Lys Val Gly Ile ValGly Arg Thr Gly Ala Gly Lys Ser Ser 1070 1075 1080 ctc atc tca gcc cttttt aga ttg tca gaa ccc gaa ggt aaa att 3294 Leu Ile Ser Ala Leu Phe ArgLeu Ser Glu Pro Glu Gly Lys Ile 1085 1090 1095 tgg att gat aag atc ttgaca act gaa att gga ctt cac gat tta 3339 Trp Ile Asp Lys Ile Leu Thr ThrGlu Ile Gly Leu His Asp Leu 1100 1105 1110 agg aag aag atg tca atc atacct cag gaa cct gtt ttg ttc act 3384 Arg Lys Lys Met Ser Ile Ile Pro GlnGlu Pro Val Leu Phe Thr 1115 1120 1125 gga aca atg agg aaa aac ctg gatccc ttt aat gag cac acg gat 3429 Gly Thr Met Arg Lys Asn Leu Asp Pro PheAsn Glu His Thr Asp 1130 1135 1140 gag gaa ctg tgg aat gcc tta caa gaggta caa ctt aaa gaa acc 3474 Glu Glu Leu Trp Asn Ala Leu Gln Glu Val GlnLeu Lys Glu Thr 1145 1150 1155 att gaa gat ctt cct ggt aaa atg gat actgaa tta gca gaa tca 3519 Ile Glu Asp Leu Pro Gly Lys Met Asp Thr Glu LeuAla Glu Ser 1160 1165 1170 gga tcc aat ttt agt gtt gga caa aga caa ctggtg tgc ctt gcc 3564 Gly Ser Asn Phe Ser Val Gly Gln Arg Gln Leu Val CysLeu Ala 1175 1180 1185 agg gca att ctc agg aaa aat cag ata ttg att attgat gaa gcg 3609 Arg Ala Ile Leu Arg Lys Asn Gln Ile Leu Ile Ile Asp GluAla 1190 1195 1200 acg gca aat gtg gat cca aga act gat gag tta ata caaaaa aaa 3654 Thr Ala Asn Val Asp Pro Arg Thr Asp Glu Leu Ile Gln Lys Lys1205 1210 1215 atc cgg gag aaa ttt gcc cac tgc acc gtg cta acc att gcacac 3699 Ile Arg Glu Lys Phe Ala His Cys Thr Val Leu Thr Ile Ala His1220 1225 1230 aga ttg aac acc att att gac agc gac aag ata atg gtt ttagat 3744 Arg Leu Asn Thr Ile Ile Asp Ser Asp Lys Ile Met Val Leu Asp1235 1240 1245 tca gga aga ctg aaa gaa tat gat gag ccg tat gtt ttg ctgcaa 3789 Ser Gly Arg Leu Lys Glu Tyr Asp Glu Pro Tyr Val Leu Leu Gln1250 1255 1260 aat aaa gag agc cta ttt tac aag atg gtg caa caa ctg ggcaag 3834 Asn Lys Glu Ser Leu Phe Tyr Lys Met Val Gln Gln Leu Gly Lys1265 1270 1275 gca gaa gcc gct gcc ctc act gaa aca gca aaa cag gta tacttc 3879 Ala Glu Ala Ala Ala Leu Thr Glu Thr Ala Lys Gln Val Tyr Phe1280 1285 1290 aaa aga aat tat cca cat att ggt cac act gac cac atg gttaca 3924 Lys Arg Asn Tyr Pro His Ile Gly His Thr Asp His Met Val Thr1295 1300 1305 aac act tcc aat gga cag ccc tcg acc tta act att ttc gag3966 Asn Thr Ser Asn Gly Gln Pro Ser Thr Leu Thr Ile Phe Glu 1310 13151320 51 1322 PRT Homo sapiens 51 Met Leu Pro Val Tyr Gln Glu Val Lys ProAsn Pro Leu Gln Asp Ala 1 5 10 15 Asn Leu Cys Ser Arg Val Phe Phe TrpTrp Leu Asn Pro Leu Phe Lys 20 25 30 Ile Gly His Lys Arg Arg Leu Glu GluAsp Asp Met Tyr Ser Val Leu 35 40 45 Pro Glu Asp Arg Ser Gln His Leu GlyGlu Glu Leu Gln Gly Phe Trp 50 55 60 Asp Lys Glu Val Leu Arg Ala Glu AsnAsp Ala Gln Lys Pro Ser Leu 65 70 75 80 Thr Arg Ala Ile Ile Lys Cys TyrTrp Lys Ser Tyr Leu Val Leu Gly 85 90 95 Ile Phe Thr Leu Ile Glu Glu SerAla Lys Val Ile Gln Pro Ile Phe 100 105 110 Leu Gly Lys Ile Ile Asn TyrPhe Glu Asn Tyr Asp Pro Met Asp Ser 115 120 125 Val Ala Leu Asn Thr AlaTyr Ala Tyr Ala Thr Val Leu Thr Phe Cys 130 135 140 Thr Leu Ile Leu AlaIle Leu His His Leu Tyr Phe Tyr His Val Gln 145 150 155 160 Cys Ala GlyMet Arg Leu Arg Val Ala Met Cys His Met Ile Tyr Arg 165 170 175 Lys AlaLeu Arg Leu Ser Asn Met Ala Met Gly Lys Thr Thr Thr Gly 180 185 190 GlnIle Val Asn Leu Leu Ser Asn Asp Val Asn Lys Phe Asp Gln Val 195 200 205Thr Val Phe Leu His Phe Leu Trp Ala Gly Pro Leu Gln Ala Ile Ala 210 215220 Val Thr Ala Leu Leu Trp Met Glu Ile Gly Ile Ser Cys Leu Ala Gly 225230 235 240 Met Ala Val Leu Ile Ile Leu Leu Pro Leu Gln Ser Cys Phe GlyLys 245 250 255 Leu Phe Ser Ser Leu Arg Ser Lys Thr Ala Thr Phe Thr AspAla Arg 260 265 270 Ile Arg Thr Met Asn Glu Val Ile Thr Gly Ile Arg IleIle Lys Met 275 280 285 Tyr Ala Trp Glu Lys Ser Phe Ser Asn Leu Ile ThrAsn Leu Arg Lys 290 295 300 Lys Glu Ile Ser Lys Ile Leu Arg Ser Ser CysLeu Arg Gly Met Asn 305 310 315 320 Leu Ala Ser Phe Phe Ser Ala Ser LysIle Ile Val Phe Val Thr Phe 325 330 335 Thr Thr Tyr Val Leu Leu Gly SerVal Ile Thr Ala Ser Arg Val Phe 340 345 350 Val Ala Val Thr Leu Tyr GlyAla Val Arg Leu Thr Val Thr Leu Phe 355 360 365 Phe Pro Ser Ala Ile GluArg Val Ser Glu Ala Ile Val Ser Ile Arg 370 375 380 Arg Ile Gln Thr PheLeu Leu Leu Asp Glu Ile Ser Gln Arg Asn Arg 385 390 395 400 Gln Leu ProSer Asp Gly Lys Lys Met Val His Val Gln Asp Phe Thr 405 410 415 Ala PheTrp Asp Lys Ala Ser Glu Thr Pro Thr Leu Gln Gly Leu Ser 420 425 430 PheThr Val Arg Pro Gly Glu Leu Leu Ala Val Val Gly Pro Val Gly 435 440 445Ala Gly Lys Ser Ser Leu Leu Ser Ala Val Leu Gly Glu Leu Ala Pro 450 455460 Ser His Gly Leu Val Ser Val His Gly Arg Ile Ala Tyr Val Ser Gln 465470 475 480 Gln Pro Trp Val Phe Ser Gly Thr Leu Arg Ser Asn Ile Leu PheGly 485 490 495 Lys Lys Tyr Glu Lys Glu Arg Tyr Glu Lys Val Ile Lys AlaCys Ala 500 505 510 Leu Lys Lys Asp Leu Gln Leu Leu Glu Asp Gly Asp LeuThr Val Ile 515 520 525 Gly Asp Arg Gly Thr Thr Leu Ser Gly Gly Gln LysAla Arg Val Asn 530 535 540 Leu Ala Arg Ala Val Tyr Gln Asp Ala Asp IleTyr Leu Leu Asp Asp 545 550 555 560 Pro Leu Ser Ala Val Asp Ala Glu ValSer Arg His Leu Phe Glu Leu 565 570 575 Cys Ile Cys Gln Ile Leu His GluLys Ile Thr Ile Leu Val Thr His 580 585 590 Gln Leu Gln Tyr Leu Lys AlaAla Ser Gln Ile Leu Ile Leu Lys Asp 595 600 605 Gly Lys Met Val Gln LysGly Thr Tyr Thr Glu Phe Leu Lys Ser Gly 610 615 620 Ile Asp Phe Gly SerLeu Leu Lys Lys Asp Asn Glu Glu Ser Glu Gln 625 630 635 640 Pro Pro ValPro Gly Thr Pro Thr Leu Arg Asn Arg Thr Phe Ser Glu 645 650 655 Ser SerVal Trp Ser Gln Gln Ser Ser Arg Pro Ser Leu Lys Asp Gly 660 665 670 AlaLeu Glu Ser Gln Asp Thr Glu Asn Val Pro Val Thr Leu Ser Glu 675 680 685Glu Asn Arg Ser Glu Gly Lys Val Gly Phe Gln Ala Tyr Lys Asn Tyr 690 695700 Phe Arg Ala Gly Ala His Trp Ile Val Phe Ile Phe Leu Ile Leu Leu 705710 715 720 Asn Thr Ala Ala Gln Val Ala Tyr Val Leu Gln Asp Trp Trp LeuSer 725 730 735 Tyr Trp Ala Asn Lys Gln Ser Met Leu Asn Val Thr Val AsnGly Gly 740 745 750 Gly Asn Val Thr Glu Lys Leu Asp Leu Asn Trp Tyr LeuGly Ile Tyr 755 760 765 Ser Gly Leu Thr Val Ala Thr Val Leu Phe Gly IleAla Arg Ser Leu 770 775 780 Leu Val Phe Tyr Val Leu Val Asn Ser Ser GlnThr Leu His Asn Lys 785 790 795 800 Met Phe Glu Ser Ile Leu Lys Ala ProVal Leu Phe Phe Asp Arg Asn 805 810 815 Pro Ile Gly Arg Ile Leu Asn ArgPhe Ser Lys Asp Ile Gly His Leu 820 825 830 Asp Asp Leu Leu Pro Leu ThrPhe Leu Asp Phe Ile Gln Thr Leu Leu 835 840 845 Gln Val Val Gly Val ValSer Val Ala Val Ala Val Ile Pro Trp Ile 850 855 860 Ala Ile Pro Leu ValPro Leu Gly Ile Ile Phe Ile Phe Leu Arg Arg 865 870 875 880 Tyr Phe LeuGlu Thr Ser Arg Asp Val Lys Arg Leu Glu Ser Thr Thr 885 890 895 Arg SerPro Val Phe Ser His Leu Ser Ser Ser Leu Gln Gly Leu Trp 900 905 910 ThrIle Arg Ala Tyr Lys Ala Glu Glu Arg Cys Gln Glu Leu Phe Asp 915 920 925Ala His Gln Asp Leu His Ser Glu Ala Trp Phe Leu Phe Leu Thr Thr 930 935940 Ser Arg Trp Phe Ala Val Arg Leu Asp Ala Ile Cys Ala Met Phe Val 945950 955 960 Ile Ile Val Ala Phe Gly Ser Leu Ile Leu Ala Lys Thr Leu AspAla 965 970 975 Gly Gln Val Gly Leu Ala Leu Ser Tyr Ala Leu Thr Leu MetGly Met 980 985 990 Phe Gln Trp Cys Val Arg Gln Ser Ala Glu Val Glu AsnMet Met Ile 995 1000 1005 Ser Val Glu Arg Val Ile Glu Tyr Thr Asp LeuGlu Lys Glu Ala 1010 1015 1020 Pro Trp Glu Tyr Gln Lys Arg Pro Pro ProAla Trp Pro His Glu 1025 1030 1035 Gly Val Ile Ile Phe Asp Asn Val AsnPhe Met Tyr Ser Pro Gly 1040 1045 1050 Gly Pro Leu Val Leu Lys His LeuThr Ala Leu Ile Lys Ser Gln 1055 1060 1065 Glu Lys Val Gly Ile Val GlyArg Thr Gly Ala Gly Lys Ser Ser 1070 1075 1080 Leu Ile Ser Ala Leu PheArg Leu Ser Glu Pro Glu Gly Lys Ile 1085 1090 1095 Trp Ile Asp Lys IleLeu Thr Thr Glu Ile Gly Leu His Asp Leu 1100 1105 1110 Arg Lys Lys MetSer Ile Ile Pro Gln Glu Pro Val Leu Phe Thr 1115 1120 1125 Gly Thr MetArg Lys Asn Leu Asp Pro Phe Asn Glu His Thr Asp 1130 1135 1140 Glu GluLeu Trp Asn Ala Leu Gln Glu Val Gln Leu Lys Glu Thr 1145 1150 1155 IleGlu Asp Leu Pro Gly Lys Met Asp Thr Glu Leu Ala Glu Ser 1160 1165 1170Gly Ser Asn Phe Ser Val Gly Gln Arg Gln Leu Val Cys Leu Ala 1175 11801185 Arg Ala Ile Leu Arg Lys Asn Gln Ile Leu Ile Ile Asp Glu Ala 11901195 1200 Thr Ala Asn Val Asp Pro Arg Thr Asp Glu Leu Ile Gln Lys Lys1205 1210 1215 Ile Arg Glu Lys Phe Ala His Cys Thr Val Leu Thr Ile AlaHis 1220 1225 1230 Arg Leu Asn Thr Ile Ile Asp Ser Asp Lys Ile Met ValLeu Asp 1235 1240 1245 Ser Gly Arg Leu Lys Glu Tyr Asp Glu Pro Tyr ValLeu Leu Gln 1250 1255 1260 Asn Lys Glu Ser Leu Phe Tyr Lys Met Val GlnGln Leu Gly Lys 1265 1270 1275 Ala Glu Ala Ala Ala Leu Thr Glu Thr AlaLys Gln Val Tyr Phe 1280 1285 1290 Lys Arg Asn Tyr Pro His Ile Gly HisThr Asp His Met Val Thr 1295 1300 1305 Asn Thr Ser Asn Gly Gln Pro SerThr Leu Thr Ile Phe Glu 1310 1315 1320 52 3 PRT Artificial SequenceDescription of Artificial Sequence T-K-F peptide motif variant 52 ThrLys Phe 1 53 3 PRT Artificial Sequence Description of ArtificialSequence T-K-F peptide motif variant 53 Thr Ala Phe 1 54 3 PRTArtificial Sequence Description of Artificial Sequence T-K-F peptidemotif variant 54 Thr Ala Leu 1 55 3 PRT Artificial Sequence Descriptionof Artificial Sequence T-K-F peptide motif variant 55 Thr Glu Leu 1 56 3PRT Artificial Sequence Description of Artificial Sequence T-K-F peptidemotif variant 56 Thr Lys Arg 1 57 3 PRT Artificial Sequence Descriptionof Artificial Sequence T-K-F peptide motif variant 57 Thr Gln Asn 1 58 3PRT Artificial Sequence Description of Artificial Sequence T-K-F peptidemotif variant 58 Ala Lys Arg 1 59 31 DNA Artificial Sequence Descriptionof Artificial Sequence oligonucleotide primer for amplifying modifiedhuman MDR3 59 agcgctagcg atggatcttg aggcggcaaa g 31 60 27 DNA ArtificialSequence Description of Artificial Sequence oligonucleotide primer foramplifying modified human MDR3 60 tacggtaccg gtgccccagc ctggaca 27 61 31DNA Artificial Sequence Description of Artificial Sequenceoligonucleotide primer for amplifying modified human MRP4 61 agcgctagcgatgctgcccg tgtaccagga g 31 62 27 DNA Artificial Sequence Description ofArtificial Sequence oligonucleotide primer for amplifying modified humanMRP4 62 tacggtaccg gtgcctcgaa aatagtt 27

We claim:
 1. A modified ABC transporter polypeptide that is localizedpredominantly in the basolateral membrane of a polarized cell or in theplasma membrane of a non-polarized cell, said modified ABC transporterpolypeptide consisting of the amino acid sequence of a native ABCtransporter polypeptide that normally accumulates in the apical(canalicular) membrane of a polarized cell wherein one or more aminoacid residues of a C-terminal tripeptide T-K-F motif of said native ABCtransporter polypeptide having a sequence set forth in any one of SEQ IDNOs: 52 to 58 is substituted or deleted.
 2. The modified ABC transporterpolypeptide of claim 1, wherein the threonine at the first position ofthe tripeptide T-K-F motif is substituted with a different amino acidresidue.
 3. The modified ABC transporter of claim 2 wherein thedifferent amino acid residue is alanine.
 4. The modified ABC transporterpolypeptide of claim 1, wherein the lysine at the second position of thetripeptide T-K-F motif is substituted with a different amino acidresidue.
 5. The modified ABC transporter polypeptide of claim 4, whereinthe different amino acid residue is alanine or proline.
 6. The modifiedABC transporter polypeptide of claim 1, wherein three amino acidresidues of the tripeptide T-K-F motif are substituted with differentamino acid residues.
 7. The modified ABC transporter polypeptide ofclaim 6, wherein each of the three amino acid residues of the tripeptideT-K-F motif is substituted with alanine.
 8. The modified ABC transporterpolypeptide of claim 6, wherein the three amino acid residues of thetripeptide T-K-F motif are substituted respectively with alanine,proline and valine.
 9. The modified ABC transporter polypeptide of claim1, wherein three amino acid residues of the tripeptide T-K-F motif aredeleted.
 10. The modified ABC transporter polypeptide of claim 1 whereinthe native ABC transporter polypeptide that normally accumulates in theapical (canalicular) membrane of a polarized cell is selected from thegroup consisting of canalicular multispecific organic anion transporter(cMOAT), MDR3 and MRP4.
 11. The modified ABC transporter polypeptide ofclaim 10, wherein the native ABC transporter polypeptide is human cMOAT,human MDR3, or human MRP4.
 12. A fusion polypeptide comprising themodified ABC transporter of claim 1 covalently linked to greenfluorescent protein (gfp).
 13. A modified canalicular multispecificorganic anion transporter (cMOAT) polypeptide localized predominantly inthe basolateral membrane of a polarized cell or in the plasma membraneof a non-polarized cell, said modified cMOAT polypeptide consisting ofthe amino acid sequence of a native cMOAT polypeptide wherein one ormore amino acid residues of a C-terminal tripeptide T-K-F motif of saidnative cMOAT polypeptide having a sequence set forth in SEQ ID NO: 52 orSEQ ID NO: 53 or SEQ ID NO: 55 is substituted or deleted.
 14. Themodified cMOAT polypeptide of claim 13 wherein one or more amino acidresidues of the C-terminal tripeptide T-K-F motif of said native cMOATpolypeptide having the sequence set forth in SEQ ID NO: 52 issubstituted or deleted.
 15. The modified cMOAT polypeptide of claim 14,wherein the threonine at the first position of the tripeptide T-K-Fmotif is substituted with a different amino acid residue.
 16. Themodified cMOAT of claim 15 wherein the different amino acid residue isalanine.
 17. The modified cMOAT polypeptide of claim 14 wherein thelysine at the second position of the tripeptide T-K-F motif issubstituted with a different amino acid residue.
 18. The modified cMOATpolypeptide of claim 17 wherein the different amino acid residue isalanine or proline.
 19. The modified cMOAT polypeptide of claim 14,wherein three amino acid residues of the tripeptide T-K-F motif aresubstituted with different amino acid residues.
 20. The modified cMOATpolypeptide of claim 19, wherein each of the three amino acid residuesof the tripeptide T-K-F motif is substituted with alanine.
 21. Themodified cMOAT polypeptide of claim 19, wherein the three amino acidresidues of the tripeptide T-K-F motif are substituted respectively withalanine, proline and valine.
 22. The modified cMOAT polypeptide of claim13, wherein three amino acid residues of the tripeptide T-K-F motif aredeleted.
 23. A fusion polypeptide comprising the modified cMOAT of claim13 covalently linked to green fluorescent protein (gfp).
 24. A modifiedMDR3 polypeptide localized predominantly in the basolateral membrane ofa polarized cell or in the plasma membrane of a non-polarized cell, saidmodified MDR3 polypeptide consisting of the amino acid sequence of anative MDR3 polypeptide wherein the C-terminal tripeptide T-K-F motif ofsaid native MDR3 polypeptide having the sequence set forth in SEQ ID NO:57 is deleted.
 25. A fusion polypeptide comprising the modified MDR3 ofclaim 24 covalently linked to green fluorescent protein (gfp).
 26. Amodified MRP4 polypeptide localized predominantly in the basolateralmembrane of a polarized cell or in the plasma membrane of anon-polarized cell, said modified MRP4 polypeptide consisting of theamino acid sequence of a native MRP4 polypeptide wherein the C-terminaltripeptide T-K-F motif of said native MRP4 polypeptide having thesequence set forth in SEQ ID NO: 54 is deleted.
 27. A fusion polypeptidecomprising the modified MRP4 of claim 26 covalently linked to greenfluorescent protein (gfp).
 28. A modified canalicular multispecificorganic anion transporter (cMOAT) polypeptide that consists of the aminoacid sequence set forth in SEQ ID NO:
 4. 29. A fusion polypeptidecomprising the modified cMOAT polypeptide of claim 28 covalently linkedto green fluorescent protein (gfp).
 30. A modified canalicularmultispecific organic anion transporter (cMOAT) polypeptide thatcomprises the amino acid sequence set forth in SEQ ID NO:
 6. 31. Afusion polypeptide comprising the modified cMOAT polypeptide of claim 30covalently linked to green fluorescent protein (gfp).
 32. A modifiedcanalicular multispecific organic anion transporter (cMOAT) polypeptidethat comprises the amino acid sequence set forth in SEQ ID NO:
 10. 33. Afusion polypeptide comprising the modified cMOAT polypeptide of claim 32covalently linked to green fluorescent protein (gfp).
 34. A modifiedcanalicular multispecific organic anion transporter (cMOAT) polypeptidethat comprises the amino acid sequence set forth in SEQ ID NO:
 12. 35. Afusion polypeptide comprising the modified cMOAT polypeptide of claim 34covalently linked to green fluorescent protein (gfp).
 36. A modifiedcanalicular multispecific organic anion transporter (cMOAT) polypeptidethat comprises the amino acid sequence set forth in SEQ ID NO:
 16. 37. Afusion polypeptide comprising the modified cMOAT polypeptide of claim 36covalently linked to green fluorescent protein (gfp).
 38. A modifiedMDR3 polypeptide that consists of the amino acid sequence set forth inSEQ ID NO:
 49. 39. A fusion polypeptide comprising the modified MDR3polypeptide of claim 38 covalently linked to green fluorescent protein(gfp).
 40. A modified MRP4 polypeptide that consists of the amino acidsequence set forth in SEQ ID NO:
 51. 41. A fusion polypeptide comprisingthe modified MRP4 polypeptide of claim 40 covalently linked to greenfluorescent protein (gfp).
 42. An isolated nucleic acid that comprises anucleotide sequence encoding a modified ABC transporter polypeptide thatconsists of the amino acid sequence of a native ABC transporterpolypeptide that normally accumulates in the apical (canalicular)membrane of a polarized cell wherein one or more amino acid residues ofa C-terminal tripeptide T-K-F motif of said native ABC transporterpolypeptide having an amino acid sequence set forth in any one of SEQ IDNOs: 52 to 58 is substituted or deleted.
 43. The isolated nucleic acidof claim 42 wherein said nucleotide sequence includes a mutationrelative to the corresponding wild-type gene selected from the groupconsisting of: (i) a deletion of nucleotides from the 3′ end of thecoding region of said wild-type gene sufficient to encode a modified ABCtransporter polypeptide lacking said T-K-F motif; (ii) a substitution ofnucleotides within the 3-end of the coding region of said wild-type genesufficient to encode a modified ABC transporter wherein the first aminoacid residue of said T-K-F motif is substituted; (iii) a substitution ofnucleotides within the 3′-end of the coding region of said wild-typegene sufficient to encode a modified ABC transporter wherein the secondamino acid residue of said T-K-F motif is substituted; and (iv) asubstitution of nucleotides within the 3′-end of the coding region ofsaid wild-type gene sufficient to encode a modified ABC transporterwherein all three amino acid residues of said T-K-F motif aresubstituted.
 44. The isolated nucleic acid of claim 43 wherein themodified ABC transporter comprises the substitution of threonine foralanine at the first amino acid residue of said T-K-F motif.
 45. Theisolated nucleic acid of claim 43 wherein the modified ABC transportercomprises the substitution of lysine for alanine or proline at thesecond amino acid residue of said T-K-F motif.
 46. The isolated nucleicacid of claim 43 wherein the modified ABC transporter comprises thesubstitution of each of the three amino acid residues of the tripeptideT-K-F motif for alanine.
 47. The isolated nucleic acid of claim 43wherein the modified ABC transporter comprises the substitution of thethree amino acid residues of the tripeptide T-K-F motif respectively foralanine, proline and valine.
 48. The isolated nucleic acid of claim 43encoding a modified canalicular multispecific organic anion transporter(cMOAT), modified MDR3 or modified MRP4 polypeptide.
 49. The isolatednucleic acid of claim 48 wherein the modified cMOAT, modified MDR3 ormodified MRP4 polypeptide is modified human cMOAT, modified human MDR3,or modified human MRP4.
 50. An isolated nucleic acid encoding a modifiedcanalicular multispecific organic anion transporter (cMOAT) polypeptide,said nucleic acid consisting of a nucleotide sequence selected from thegroup consisting of: (i) the nucleotide sequence set forth in SEQ ID NO:3; and (ii) a nucleotide sequence encoding the amino acid sequence setforth in SEQ ID NO:
 4. 51. An isolated nucleic acid encoding a modifiedcanalicular multispecific organic anion transporter (cMOAT) polypeptide,said nucleic acid comprising a nucleotide sequence selected from thegroup consisting of (i) the nucleotide sequence set forth in SEQ ID NO:5; and (ii) a nucleotide sequence encoding the amino acid sequence setforth in SEQ ID NO:
 6. 52. An isolated nucleic acid encoding a modifiedcanalicular multispecific organic anion transporter (cMOAT) polypeptide,said nucleic acid comprising a nucleotide sequence selected from thegroup consisting of: (i) the nucleotide sequence set forth in SEQ ID NO:9; and (ii) a nucleotide sequence encoding the amino acid sequence setforth in SEQ ID NO:
 10. 53. An isolated nucleic acid encoding a modifiedcanalicular multispecific organic anion transporter (cMOAT) polypeptide,said nucleic acid comprising a nucleotide sequence selected from thegroup consisting of: (i) the nucleotide sequence set forth in SEQ ID NO:11; and (iii) a nucleotide sequence encoding the amino acid sequence setforth in SEQ ID NO:
 12. 54. An isolated nucleic acid encoding a modifiedcanalicular multispecific organic anion transporter (cMOAT) polypeptide,said nucleic acid comprising a nucleotide sequence selected from thegroup consisting of: (i) the nucleotide sequence set forth in SEQ ID NO:15; and (ii) a nucleotide sequence encoding the amino acid sequence setforth in SEQ ID NO:
 16. 55. An isolated nucleic acid encoding a modifiedMDR3 polypeptide, said nucleic acid consisting of a nucleotide sequenceselected from the group consisting of: (i) the nucleotide sequence setforth in SEQ ID NO: 48; and (ii) a nucleotide sequence encoding theamino acid sequence set forth in SEQ ID NO:
 49. 56. An isolated nucleicacid encoding a modified MRP4 polypeptide, said nucleic acid consistingof a nucleotide sequence selected from the group consisting of: (i) thenucleotide sequence set forth in SEQ ID NO: 50; and (ii) a nucleotidesequence encoding the amino acid sequence set forth in SEQ ID NO: 51.57. A method of producing nucleic acid encoding a modified ABCtransporter polypeptide comprising deleting or substituting a portion ofthe coding region of nucleic acid encoding an ABC transporterpolypeptide that accumulates in the apical (canalicular) membrane of apolarized cell, said portion encoding the first or second amino acidresidue of the C-terminal tripeptide T-K-F motif of said ABC transporteror all three amino acid residues of said C-terminal tripeptide T-K-Fmotif, wherein said motif has a sequence set forth in any one of SEQ IDNOs: 52 to
 58. 58. A method of producing nucleic acid encoding amodified human canalicular multispecific organic anion transporter(cMOAT) polypeptide comprising deleting or substituting a portion of thecoding region set forth in SEQ ID NO: 1, said portion encoding the firstor second amino acid residue of the C-terminal tripeptide T-K-F motif ofsaid cMOAT or all three amino acid residues of said C-terminaltripeptide T-K-F motif, wherein said motif has a sequence set forth inSEQ ID NO:
 52. 59. The method of claim 58 wherein deleting orsubstituting a portion of the coding region comprises amplifying nucleicacid encoding cMOAT using a primer comprising a deletion or substitutionwithin the sequence corresponding or complementary to the portion of thecoding region encoding the first or second amino acid residue of theC-terminal tripeptide T-K-F motif of said cMOAT or all three amino acidresidues of said C-terminal tripeptide T-K-F motif; and selecting theamplified nucleic acid wherein the deletion or substitution isintroduced into the remainder of the coding region of SEQ ID NO:
 1. 60.The method of claim 58 wherein the primer comprises or is complementaryto a nucleotide sequence selected from the group consisting of SEQ IDNO: 26; SEQ ID NO: 27; SEQ ID NO: 29; SEQ ID NO: 30; SEQ ID NO: 32; andSEQ ID NO:
 33. 61. A method of producing nucleic acid encoding amodified human MDR3 polypeptide comprising deleting a portion of thecoding region encoding a native human MDR3 polypeptide, said portionencoding the C-terminal tripeptide T-K-F motif of said native humanMDR3, wherein said motif has a sequence set forth in SEQ ID NO:
 57. 62.The method of claim 61 wherein deleting a portion of the coding regioncomprises amplifying nucleic acid encoding native human MDR3 using aprimer that hybridizes to the 3′-end of said coding region or itscomplement wherein said primer comprises a deletion within the sequencecorresponding or complementary to the portion of said coding regionencoding said C-terminal tripeptide T-K-F motif; and selecting theamplified nucleic acid wherein the deletion is introduced into theremainder of the coding region encoding MDR3.
 63. The method of claim 62wherein the primer comprises or is complementary to the nucleotidesequence set forth in SEQ ID NO:
 60. 64. A method of producing nucleicacid encoding a modified human MRP4 polypeptide comprising deleting aportion of the coding region encoding a native human MRP4 polypeptide,said portion encoding the C-terminal tripeptide T-K-F motif of saidnative human MRP4, wherein said motif has a sequence set forth in SEQ IDNO:
 54. 65. The method of claim 64 wherein deleting a portion of thecoding region comprises amplifying nucleic acid encoding native humanMRP4 using a primer that hybridizes to the 3′-end of said coding regionor its complement wherein said primer comprises a deletion within thesequence corresponding or complementary to the portion of said codingregion encoding said C-terminal tripeptide T-K-F motif; and selectingthe amplified nucleic acid wherein the deletion is introduced into theremainder of the coding region encoding MRP4.
 66. The method of claim 65wherein the primer comprises or is complementary to the nucleotidesequence set forth in SEQ ID NO:
 62. 67. A gene construct comprising theisolated nucleic acid of claim 42 encoding a modified ABC transporterpolypeptide wherein said nucleic acid is in operable connection with apromoter sequence to facilitate expression of said polypeptide in acell.
 68. The gene construct of claim 67 wherein the nucleic acid is inthe same reading frame as nucleic acid encoding green fluorescentprotein (gfp) such that the modified ABC transporter polypeptide iscapable of being expressed as a fusion polypeptide with said gfp.
 69. Agene construct comprising the isolated nucleic acid of claim 50 encodinga modified ABC transporter polypeptide wherein said nucleic acid is inoperable connection with a promoter sequence to facilitate expression ofsaid polypeptide in a cell.
 70. The gene construct of claim 69 whereinthe nucleic acid is in the same reading frame as nucleic acid encodinggreen fluorescent protein (gfp) such that the modified ABC transporterpolypeptide is capable of being expressed as a fusion polypeptide withsaid gfp.
 71. A gene construct comprising the isolated nucleic acid ofclaim 51 encoding a modified ABC transporter polypeptide wherein saidnucleic acid is in operable connection with a promoter sequence tofacilitate expression of said polypeptide in a cell.
 72. The geneconstruct of claim 71 wherein the nucleic acid is in the same readingframe as nucleic acid encoding green fluorescent protein (gfp) such thatthe modified ABC transporter polypeptide is capable of being expressedas a fusion polypeptide with said gfp.
 73. A gene construct comprisingthe isolated nucleic acid of claim 52 encoding a modified ABCtransporter polypeptide wherein said nucleic acid is in operableconnection with a promoter sequence to facilitate expression of saidpolypeptide in a cell.
 74. The gene construct of claim 73 wherein thenucleic acid is in the same reading frame as nucleic acid encoding greenfluorescent protein (gfp) such that the modified ABC transporterpolypeptide is capable of being expressed as a fusion polypeptide withsaid gfp.
 75. A gene construct comprising the isolated nucleic acid ofclaim 53 encoding a modified ABC transporter polypeptide wherein saidnucleic acid is in operable connection with a promoter sequence tofacilitate expression of said polypeptide in a cell.
 76. The geneconstruct of claim 75 wherein the nucleic acid is in the same readingframe as nucleic acid encoding green fluorescent protein (gfp) such thatthe modified ABC transporter polypeptide is capable of being expressedas a fusion polypeptide with said gfp.
 77. A gene construct comprisingthe isolated nucleic acid of claim 54 encoding a modified ABCtransporter polypeptide wherein said nucleic acid is in operableconnection with a promoter sequence to facilitate expression of saidpolypeptide in a cell.
 78. The gene construct of claim 77 wherein thenucleic acid is in the same reading frame as nucleic acid encoding greenfluorescent protein (gfp) such that the modified ABC transporterpolypeptide is capable of being expressed as a fusion polypeptide withsaid gfp.
 79. A gene construct comprising the isolated nucleic acid ofclaim 55 encoding a modified ABC transporter polypeptide wherein saidnucleic acid is in operable connection with a promoter sequence tofacilitate expression of said polypeptide in a cell.
 80. The geneconstruct of claim 79 wherein the nucleic acid is in the same readingframe as nucleic acid encoding green fluorescent protein (gfp) such thatthe modified ABC transporter polypeptide is capable of being expressedas a fusion polypeptide with said gfp.
 81. A gene construct comprisingthe isolated nucleic acid of claim 56 encoding a modified ABCtransporter polypeptide wherein said nucleic acid is in operableconnection with a promoter sequence to facilitate expression of saidpolypeptide in a cell.
 82. The gene construct of claim 81 wherein thenucleic acid is in the same reading frame as nucleic acid encoding greenfluorescent protein (gfp) such that the modified ABC transporterpolypeptide is capable of being expressed as a fusion polypeptide withsaid gfp.
 83. A method of enhancing the resistance of a cell to one ormore chemical compounds comprising expressing a modified ABC transporterpolypeptide in said cell for a time and under conditions sufficient forsaid cell to have modified growth and/or viability in the presence ofsaid compound, wherein said modified ABC transporter comprises the aminoacid sequence of the corresponding native ABC transporter wherein one ormore amino acid residues of a C-terminal tripeptide T-K-F motif of saidnative ABC transporter polypeptide having a sequence set forth in anyone of SEQ ID NOs: 52 to 58 is substituted or deleted.
 84. The method ofclaim 83 wherein the native ABC transporter is selected from the groupconsisting of canalicular multispecific organic anion transporter(cMOAT), MDR3 and MRP4.
 85. The method of claim 83 wherein said cell isa polarized cell and wherein said modified ABC transporter is localizedpredominantly in the basolateral membrane of said polarized cell. 86.The method of claim 85 wherein the polarized cell is an epithelial cell.87. The method of claim 86 wherein the epithelial cell is selected fromthe group consisting of: a cultured MDCK cell, a cultured Caco-2 cell, ahepatocyte, an intestinal cell, and a hippocampal neuron.
 88. The methodof claim 83 wherein said cell is a non-polarized cell and wherein saidmodified ABC transporter is localized in the plasma membrane of saidnon-polarized cell.
 89. The method of claim 88 wherein the non-polarizedcell is selected from the group consisting of a fibroblast, ahaemopoietic cell, a cultured L1210 cell and a cultured Jurkat cell. 90.The method of claim 83 wherein the chemical compound is a cytotoxic orcytostatic compound selected from the group consisting of: (i) achemotherapeutic agent that is capable of being transported from thecell by the modified ABC transporter; (ii) an anti-bacterial compoundthat is capable of being transported from the cell by the modified ABCtransporter; and (iii) an anti-fungal compound that is capable of beingtransported from the cell by the modified ABC transporter.
 91. Themethod of claim 83 wherein the efflux of the chemical compound from thecell is enhanced by the expressed modified ABC transporter therebyenhancing resistance of the cell to said chemical compound.
 92. Themethod of claim 91 wherein the chemical compound is transported from thecell as a glutathione conjugate.
 93. The method of claim 83 furthercomprising introducing nucleic acid into the cell encoding the modifiedABC transporter.
 94. A method of enhancing the resistance of anon-polarized cell to Busulfan comprising expressing a modified cMOATpolypeptide in said cell for a time and under conditions sufficient forsaid cell to have modified growth and/or viability in the presence ofBusulfan, wherein said modified cMOAT polypeptide comprises the aminoacid sequence of native cMOAT wherein one or more amino acid residues ofthe C-terminal tripeptide T-K-F motif of said native cMOAT having asequence set forth in SEQ ID NO: 52 or SEQ ID NO: 53 or SEQ ID NO: 55 issubstituted or deleted.
 95. The method of claim 94 wherein the modifiedcMOAT polypeptide comprises the amino acid sequence of native cMOATwherein one or more amino acid residues of the C-terminal tripeptideT-K-F motif of SEQ ID NO: 52 is substituted or deleted, saidsubstitution or deletion selected from the group consisting of: (i)substitution of threonine at the first position of the tripeptide T-K-Fmotif for alanine; (ii) substitution of lysine at the second position ofthe tripeptide T-K-F motif for a different amino acid residue; (iii)substitution of all three amino acid residues of the tripeptide T-K-Fmotif for different amino acid residues; and (iv) deletion of the threeamino acid residues of the tripeptide T-K-F motif.
 96. The method ofclaim 95 wherein the different amino acid residue at (ii) is alanine orproline.
 97. The method of claim 95 wherein each different amino acidresidue at (iii) is alanine.
 98. The method of claim 95 wherein thedifferent amino acid residues at (iii) are alanine, proline and valine,respectively.
 99. The method of claim 94 further comprising introducingnucleic acid into the non-polarized cell encoding the modified cMOATpolypeptide.
 100. The method of claim 99 wherein the nucleic acidencoding the modified cMOAT polypeptide consists of the nucleotidesequence set forth in SEQ ID NO: 3 or a degenerate nucleotide sequencethereto.
 101. A method of enhancing the resistance of a non-polarizedcell to Busulfan comprising expressing a modified cMOAT polypeptide insaid cell for a time and under conditions sufficient for said cell tohave modified growth and/or viability in the presence of Busulfan,wherein said modified cMOAT polypeptide consists of the amino acidsequence set forth in SEQ ID NO:
 4. 102. The method of claim 101 furthercomprising introducing nucleic acid into the non-polarized cell encodingthe modified cMOAT polypeptide.
 103. The method of claim 102 wherein thenucleic acid encoding the modified cMOAT polypeptide consists of thenucleotide sequence set forth in SEQ ID NO: 3 or a degenerate nucleotidesequence thereto.
 104. A method of protecting a non-polarized cell of anorganism or tissue comprising said non-polarized cell during theadministration of a cytotoxic or cytostatic chemical compound to asubject, said method comprising: (i) expressing the modified ABCtransporter polypeptide of claim 1 in said non-polarized cell for a timeand under conditions sufficient for said cell to efficiently transportsaid cytotoxic or cytostatic compound from said cell or otherwiseacquire resistance to said compound; and (ii) optionally administeringan amount of an inhibitor of a native ABC transporter sufficient toablate or inhibit the growth of a cell expressing said native ABCtransporter, wherein said native ABC transporter is different to thatfrom which said modified ABC transporter polypeptide is derived and isinvolved in the transport of said cytotoxic or cytostatic chemicalcompound and wherein said cell expressing said native ABC transporter isdifferent to the non-polarized cell expressing the modified ABCtransporter.
 105. The method of claim 104 wherein the non-polarized cellexpressing the modified ABC transporter is a cell of the haematopoieticsystem and wherein the cell expressing the native ABC transporter is anepithelial cell.
 106. The method of claim 105 wherein the epithelialcell is selected from the group consisting of: hepatocyte, intestinalcell, and hippocampal neuron.
 107. The method of claim 104 wherein thecell expressing the native ABC transporter is a polarized tumor cell ornon-polarized tumor cell.
 108. The method of claim 104 wherein themodified ABC transporter at (i) is a modified cMOAT polypeptide andwherein the native ABC transporter at (ii) is selected from the groupconsisting of native MRP1, native MRP3, native MRP4, native MRP5, nativeMRP6, native MDR3, and native P-gp.
 109. The method of claim 108 themodified ABC transporter at (i) is a modified cMOAT polypeptide andwherein the native ABC transporter at (ii) is native MRP1 and whereinthe cytostatic or cytotoxic compound is a substrate of MRP1 or MRP2.110. The method of claim 108 wherein the modified ABC transporter at (i)is a modified cMOAT polypeptide and wherein the native ABC transporterat (ii) is native MRP3 and wherein the cytostatic or cytotoxic compoundis a substrate of MRP2 or MRP3.
 111. The method of claim 104 wherein themodified ABC transporter at (i) is a modified MDR3 polypeptide andwherein the native ABC transporter at (ii) is selected from the groupconsisting of native MRP1, native MRP2, native MRP3, native MRP4, nativeMRP5, native MRP6, and native P-gp.
 112. The method of claim 111 whereinthe native ABC transporter at (ii) is native MRP2 and wherein theinhibitor of native MRP2 is selected from the group consisting of:α-Naphthylisothiocyanate, Chlorpromazine, Cyclosporin,Estradiol-17β-glucuronide, Ethinylestradiol,Glycolithocholate-3α-O-sulfate, Lithocholate-3α-glucuronide,Manganese-bilirubin, Phalloidin, Taurocholate, and Taurolithocholate.113. The method of claim 104 wherein the modified ABC transporter at (i)is a modified MRP4 polypeptide and wherein the native ABC transporter at(ii) is selected from the group consisting of native MRP1, native MRP2,native MRP3, native MRP5, native MRP6, native MDR3 and native P-gp. 114.The method of claim 113 wherein the native ABC transporter at (ii) isnative MRP2 and wherein the inhibitor of native MRP2 is selected fromthe group consisting of: α-Naphthylisothiocyanate, Chlorpromazine,Cyclosporin, Estradiol-17β-glucuronide, Ethinylestradiol,Glycolithocholate-3α-O-sulfate, Lithocholate-3α-O-glucuronide,Manganese-bilirubin, Phalloidin, Taurocholate, and Taurolithocholate.115. A method of enhancing the resistance of a polarized cell of anorganism or tissue comprising said polarized cell during theadministration of a cytotoxic or cytostatic chemical compound to asubject, said method comprising: (i) expressing the modified ABCtransporter polypeptide of claim 1 in said polarized cell for a time andunder conditions sufficient for said cell to enhance transport saidcytotoxic or cytostatic compound from said cell or otherwise enhanceresistance to said compound; and (ii) optionally, administering anamount of an inhibitor of a native ABC transporter sufficient to ablateor inhibit the growth of a cell expressing said native ABC transporter,wherein said native ABC transporter is different to that from which saidmodified ABC transporter polypeptide is derived and is involved in thetransport of said cytotoxic or cytostatic chemical compound and whereinsaid cell expressing said native ABC transporter is different to thepolarized cell expressing the modified ABC transporter.
 116. The methodof claim 115 wherein the polarized cell expressing the modified ABCtransporter is an epithelial cell and wherein the cell expressing thenative ABC transporter is a non-polarized cell.
 117. The method of claim116 wherein the epithelial cell is selected from the group consistingof: hepatocyte, intestinal cell, and hippocampal neuron.
 118. The methodof claim 116 wherein the cell expressing the native ABC transporter is anon-polarized cell of the haematopoletic system.
 119. The method ofclaim 115 wherein the modified ABC transporter at (i) is a modifiedcMOAT polypeptide and wherein the native ABC transporter at (ii) isselected from the group consisting of native MRP1, native MRP3, nativeMRP4, native MRP5, native MRP6, native MDR3, and native P-gp.
 120. Themethod of claim 115 wherein the modified ABC transporter at (i) is amodified cMOAT polypeptide and wherein the native ABC transporter at(ii) is native MRP1 and wherein the cytostatic or cytotoxic compound isa substrate of MRP1 or MRP2.
 121. The method of claim 115 wherein themodified ABC transporter at (i) is a modified cMOAT polypeptide andwherein the native ABC transporter at (ii) is native MRP3 and whereinthe cytostatic or cytotoxic compound is a substrate of MRP2 or MRP3.122. The method of claim 115 wherein the modified ABC transporter at (i)is a modified MDR3 polypeptide and wherein the native ABC transporter at(ii) is selected from the group consisting of native MRP1, native MRP2,native MRP3, native MRP4, native MRP5, native MRP6, and native P-gp.123. The method of claim 122 wherein the native ABC transporter at (ii)is native MRP2 and wherein the inhibitor of native MRP2 is selected fromthe group consisting of α-Naphthylisothiocyanate, Chlorpromazine,Cyclosporin, Estradiol-17β-glucuronide, Ethinylestradiol,Glycolithocholate-3α-O-sulfate, Lithocholate-3α-O-glucuronide,Manganese-bilirubin, Phalloidin, Taurocholate, and Taurolithocholate.124. The method of claim 115 wherein the modified ABC transporter at (i)is a modified MRP4 polypeptide and wherein the native ABC transporter at(ii) is selected from the group consisting of native MRP1, native MRP2,native MRP3, native MRP5, native MRP6, native MDR3 and native P-gp. 125.The method of claim 124 wherein the native ABC transporter at (ii) isnative MRP2 and wherein the inhibitor of native MRP2 is selected fromthe group consisting of: α-Naphthylisothiocyanate, Chlorpromazine,Cyclosporin, Estradiol-17β-glucuronide, Ethinylestradiol,Glycolithocholate-3α-O-sulfate, Lithocholate-3α-O-glucuronide,Manganese-bilirubin, Phalloidin, Taurocholate, and Taurolithocholate.126. An isolated cell transformed with the gene construct of claim 67,wherein said cell expresses a modified ABC transporter polypeptide. 127.An isolated cell transformed with the gene construct of claim 68,wherein said cell expresses a modified ABC transporter polypeptide. 128.An isolated cell transformed with the gene construct of claim 69,wherein said cell expresses a modified ABC transporter polypeptide. 129.An isolated cell transformed with the gene construct of claim 70,wherein said cell expresses a modified ABC transporter polypeptide. 130.An isolated cell transformed with the gene construct of claim 71,wherein said cell expresses a modified ABC transporter polypeptide. 131.An isolated cell transformed with the gene construct of claim 72,wherein said cell expresses a modified ABC transporter polypeptide. 132.An isolated cell transformed with the gene construct of claim 73,wherein said cell expresses a modified ABC transporter polypeptide. 133.An isolated cell transformed with the gene construct of claim 74,wherein said cell expresses a modified ABC transporter polypeptide. 134.An isolated cell transformed with the gene construct of claim 75,wherein said cell expresses a modified ABC transporter polypeptide. 135.An isolated cell transformed with the gene construct of claim 76,wherein said cell expresses a modified ABC transporter polypeptide. 136.An isolated cell transformed with the gene construct of claim 77,wherein said cell expresses a modified ABC transporter polypeptide. 137.An isolated cell transformed with the gene construct of claim 78,wherein said cell expresses a modified ABC transporter polypeptide. 138.An isolated cell transformed with the gene construct of claim 79,wherein said cell expresses a modified ABC transporter polypeptide. 139.An isolated cell transformed with the gene construct of claim 80,wherein said cell expresses a modified ABC transporter polypeptide. 140.An isolated cell transformed with the gene construct of claim 81,wherein said cell expresses a modified ABC transporter polypeptide. 141.An isolated cell transformed with the gene construct of claim 82,wherein said cell expresses a modified ABC transporter polypeptide. 142.An isolated MDCK cell having a modified cMOAT polypeptide predominantlyin the basolateral membrane, said modified cMOAT polypeptide having anamino acid sequence selected from the group consisting of (1) a sequenceconsisting of SEQ ID NO: 4; (ii) a sequence comprising SEQ ID NO: 6;(iii) a sequence comprising SEQ ID NO: 10; and (iv) a sequencecomprising SEQ ID NO:
 16. 143. An isolated L1210 cell having a modifiedcMOAT polypeptide predominantly in the plasma membrane, said modifiedcMOAT polypeptide having an amino acid sequence selected from the groupconsisting of: (i) a sequence consisting of SEQ ID NO: 4; (ii) asequence comprising SEQ ID NO: 6; (iii) a sequence comprising SEQ ID NO:10; and (iv) a sequence comprising SEQ ID NO:
 16. 144. The isolatedL1210 cell of claim 143 wherein said cell has enhanced resistance toBusulfan compared to an L1210 cell not expressing said modified cMOATpolypeptide.
 145. An isolated MDCK cell having a modified MDR3polypeptide predominantly in the basolateral membrane, said modifiedMDR3 polypeptide having the amino acid sequence of SEQ ID NO:
 49. 146.An isolated L1210 cell having a modified MDR3 polypeptide predominantlyin the plasma membrane, said modified MDR3 polypeptide having the aminoacid sequence of SEQ ID NO:
 49. 147. An isolated MDCK cell having amodified MRP4 polypeptide predominantly In the basolateral membrane,said modified MRP4 polypeptide having the amino acid sequence of SEQ IDNO:
 51. 148. An isolated L1210 cell having a modified MRP4 polypeptidepredominantly in the plasma membrane, said modified MRP4 polypeptidehaving the amino acid sequence of SEQ ID NO:
 51. 149. An isolated celltransformed with nucleic acid encoding a modified ABC transporterpolypeptide, said modified ABC transporter polypeptide consisting of theamino acid sequence of a native ABC transporter polypeptide thatnormally accumulates in the apical (canalicular) membrane of a polarizedcell wherein one or more amino acid residues of a C-terminal tripeptideT-K-F motif of said native ABC transporter polypeptide having a sequenceset forth in any one of SEQ ID NOs: 52 to 58 is substituted or deleted.150. The isolated cell of claim 148 wherein said cell is a polarizedcell and wherein said modified ABC transporter accumulates predominantlyin the basolateral membrane of said cell.
 151. The isolated cell ofclaim 148, wherein said cell is a non-polarized cell and wherein saidmodified ABC transporter accumulates predominantly in the plasmamembrane of said cell.
 152. A process for identifying a substrate of anative ABC transporter polypeptide that normally accumulates in theapical (canalicular) membrane of a polarized cell, said processcomprising: (i) expressing the corresponding modified ABC transporterpolypeptide In a cell, wherein said modified ABC transporter polypeptideconsists of the amino acid sequence of said native ABC transporterpolypeptide wherein one or more amino acid residues of a C-terminalT-K-F motif of said native ABC transporter polypeptide having a sequenceset forth in any one of SEQ ID NOs: 52 to 58 is substituted or deleted;and (iii) determining the efflux of the compound from the cellexpressing the modified ABC transporter relative to a cell that does notexpress the native ABC transporter or the corresponding modified ABCtransporter, wherein efflux from the cell expressing the modified ABCtransporter indicates that the compound is a substrate for thecorresponding native ABC transporter.
 153. The process of claim 152wherein efflux is determined by measuring the amount of a conjugate ofthe compound that is exported from the cell.
 154. The process of claim153 wherein the conjugate is a glutathione conjugate.
 155. The processof claim 152 wherein the cell is a L1210 cell.
 156. The process of claim152 wherein the cell is an MDCK cell.
 157. A process for identifying anantagonist of a native ABC transporter polypeptide that normallyaccumulates in the apical (canalicular) membrane of a polarized cell,said process comprising: (i) expressing the corresponding modified ABCtransporter polypeptide in a cell, wherein said modified ABC transporterpolypeptide consists of the amino acid sequence of said native ABCtransporter polypeptide wherein one or more amino acid residues of aC-terminal T-K-F motif of said native ABC transporter polypeptide havinga sequence set forth in any one of SEQ ID NOs: 52 to 58 is substitutedor deleted; and; (ii) incubating the cell in the presence of (a) acompound being tested for its ability to antagonize activity of thenative ABC transporter polypeptide; and (b) a known substrate compoundfor said native ABC transporter polypeptide; (iii) in a separate sampleto (ii), incubating the cell in the presence of said substrate compound;and (iv) comparing the efflux of the substrate compound at (ii) and(iii), wherein reduced efflux at (ii) compared to (iii) indicates thatthe compound being tested is an antagonist of said native ABCtransporter polypeptide.
 158. The process of claim 157 wherein the knownsubstrate is selected from the group consisting of leukotriene C4(LTC4); bilirubin; monoglucuronosyl bilirubin; bisglucuronosylbilirubin; leukotriene D4 (LTD4); 1,3-chloro-2,4 dinitrobenzene;mono-chlorobimane (thiolyte); 7-chloro-4-nitrobenz-2-oxa-1,3-diazole;17β-glucuronosyl estradiol; 3α-sulfatolithocholyl taurine; Fluo-3;glutathione disulphide; p-aminohippurate; digoxin; paclitaxel;verapamil; vinblastine; phosphatidylcholine; short chainphosphatidylcholine analogue; [α-³²P]8-azido-ATP; [α-³²P]ATP;[³H]verapamil; azidothymidine monophosphate;9-(2-phosphonylmethoxyethyl)adenine (PMEA); 6-mercaptopurine; cAMP;cGMP; Sildenafil (Pfizer); Trequinsin (Sigma); and Zaprinast (Sigma).159. The process of claim 158 wherein the substrate is selected from thegroup consisting of 1-chloro-2,4-dinitrobenzene; mono-chlorobimane(thiolyte); 7-chloro-4-nitrobenz-2-oxa-1,3-diazole;); 6-mercaptopurine;and paclitaxel.
 160. The process of claim 157 wherein the cell is aL1210 cell.
 161. The process of claim 157 wherein the cell is an MDCKcell.
 162. A process for identifying an agonist of a native ABCtransporter polypeptide that normally accumulates in the apical(canalicular) membrane of a polarized cell, said process comprising: (i)expressing the corresponding modified ABC transporter polypeptide in acell, wherein said modified ABC transporter polypeptide consists of theamino acid sequence of said native ABC transporter polypeptide whereinone or more amino acid residues of a C-terminal T-K-F motif of saidnative ABC transporter polypeptide having a sequence set forth in anyone of SEQ ID NOs: 52 to 58 is substituted or deleted; and; (ii)incubating the cell in the presence of (a) a compound being tested forits ability to agonize activity of the native ABC transporterpolypeptide; and (b) a known substrate compound for said native ABCtransporter polypeptide; (iii) in a separate sample to (ii), incubatingthe cell in the presence of said substrate compound; and (iv) comparingthe efflux of the substrate compound at (ii) and (iii), wherein enhancedefflux at (ii) compared to (iii) indicates that the compound beingtested is an agonist of said native ABC transporter polypeptide. 163.The process of claim 162 wherein the known substrate is selected fromthe group consisting of: leukotriene C4 (LTC4); bilirubin;monoglucuronosyl bilirubin; bisglucuronosyl bilirubin; leukotriene D4(LTD4); 1,3-chloro-2,4-dinitrobenzene; mono-chlorobimane (thiolyte);7-chloro-4-nitrobenz-2-oxa-1,3-diazole; 17β-glucuronosyl estradiol;3α-sulfatolithocholyl taurine; Fluo-3; glutathione disulphide;p-aminohippurate; digoxin; paclitaxel; verapamil; vinblastine;phosphatidylcholine; short chain phosphatidylcholine analogue;[α-³²P]8-azido-ATP; [α-³²P]ATP; [³H]verapamil; azidothymidinemonophosphate; 9-(2-phosphonylmethoxyethyl)adenine (PMEA);6-mercaptopurine; cAMP; cGMP; Sildenafil (Pfizer); Trequinsin (Sigma);and Zaprinast (Sigma).
 164. The process of claim 163 wherein thesubstrate is selected from the group consisting of1-chloro-2,4-dinitrobenzene; mono-chlorobimane (thiolyte);7-chloro-4-nitrobenz-2-oxa-1,3-diazole;); 6-mercaptopurine; andpaclitaxel.
 165. The process of claim 162 wherein the cell is a L1210cell.
 166. The process of claim 162 wherein the cell is an MDCK cell.167. A process for identifying a modulator of a native ABC transporterpolypeptide that normally accumulates in the apical (canalicular)membrane of a polarized cell, said process comprising: (i) expressingthe corresponding modified ABC transporter polypeptide in a cell,wherein said modified ABC transporter polypeptide consists of the aminoacid sequence of said native ABC transporter polypeptide wherein one ormore amino acid residues of a C-terminal T-K-F motif of said native ABCtransporter polypeptide having a sequence set forth in any one of SEQ IDNOs: 52 to 58 is substituted or deleted; and; (ii) incubating the cellin the presence of 1-chloro-2,4-dinitrobenzene; (iii) incubating thecell at (ii) in the presence of (a) the compound being tested for itsability to modulate activity of the native ABC transporter polypeptide;and (iv) comparing the efflux of 2,4-dinitro-phenylglutathione (DNP-GS)at (ii) and (iii), wherein enhanced efflux at (ii) compared to (iii)indicates that the compound being tested is an agonist of said nativeABC transporter polypeptide and wherein reduced efflux at (ii) comparedto (iii) indicates that the compound being tested is an antagonist ofsaid native ABC transporter polypeptide.
 168. The process of claim 167wherein the cell is a L1210 cell.
 169. The process of claim 167 whereinthe cell is an MDCK cell.
 170. A process for identifying a modulator ofa native canalicular multispecific organic anion transporter (cMOAT)polypeptide, said process comprising: (i) expressing a modified cMOATpolypeptide in an L1210 cell, wherein said modified cMOAT polypeptidecomprises the amino acid sequence of native cMOAT wherein one or moreamino acid residues of the C terminal tripeptide T-K-F motif of saidnative cMOAT having a sequence set forth in SEQ ID NO: 52 or SEQ ID NO:53 or SEQ ID NO: 55 is substituted or deleted; (ii) incubating the cellin the presence of 1-chloro-2,4-dinitrobenzene; (iii) incubating thecell at (ii) in the presence of (a) the compound being tested for itsability to modulate activity of the native cMOAT polypeptide; and (iv)comparing the efflux of 2,4-dinitro-phenylglutathione (DNP-GS) at (ii)and (iii), wherein enhanced efflux at (ii) compared to (iii) indicatesthat the compound being tested is an agonist of said native cMOATpolypeptide and wherein reduced efflux at (ii) compared to (iii)indicates that the compound being tested is an antagonist of said nativecMOAT polypeptide.
 171. The process of claim 170 wherein said modifiedcMOAT polypeptide has an amino acid sequence selected from the groupconsisting of: (i) a sequence consisting of SEQ ID NO: 4; (ii) asequence comprising SEQ ID NO: 6; (iii) a sequence comprising SEQ ID NO:10; and (iv) a sequence comprising SEQ ID NO:
 16. 172. A process foridentifying a modulator of a native MDR3 polypeptide, said processcomprising: (i) expressing a modified MDR3 polypeptide in an L1210 cell,wherein said modified MDR3 polypeptide consists of the amino acidsequence of SEQ ID NO: 49; (ii) incubating the cell in the presence of[³H]paclitaxel; (iii) incubating the cell at (ii) in the presence of (a)the compound being tested for its ability to modulate activity of thenative MDR3 polypeptide; and (iv) comparing the efflux of [³H]paclitaxelat (ii) and (iii), wherein enhanced efflux at (ii) compared to (iii)indicates that the compound being tested is an agonist of said nativeMDR3 polypeptide and wherein reduced efflux at (ii) compared to (iii)indicates that the compound being tested is an antagonist of said nativeMDR3 polypeptide.
 173. A process for identifying a modulator of a nativeMRP4 polypeptide, said process comprising: (I) expressing a modifiedMRP4 polypeptide in an L1210 cell, wherein said modified MRP4polypeptide consists of the amino acid sequence of SEQ ID NO: 51; (ii)incubating the cell in the presence of 6-mercaptopurine; (iii)incubating the cell at (ii) in the presence of (a) the compound beingtested for its ability to modulate activity of the native MRP4polypeptide; and (iv) comparing the efflux of 6-thio-IMP at (ii) and(iii), wherein enhanced efflux at (ii) compared to (iii) indicates thatthe compound being tested is an agonist of said native MRP4 polypeptideand wherein reduced efflux at (ii) compared to (iii) indicates that thecompound being tested is an antagonist of said native MRP4 polypeptide.